The protective effect of resveratrol on largemouth bass (Micropterus salmoides) during out-of-season spawning.

In aquaculture production, out-of-season spawning is beneficial to solve the seasonal shortage of fry that are normally produced once annually by species such as largemouth bass (Micropterus salmoides), thereby implementing year-round fry production. Maintaining low temperature over a period of several months can delay largemouth bass ovarian development, but it can cause severe stress to their reproductive function, leading to decreased fertility during out-of-season spawning. Feeding with antioxidants is one of the most effective methods to alleviate the negative effects of low temperature stress. Therefore, the purpose of this study is to: (a) evaluate the changes in oocyte morphology, antioxidant capacity, reproductive hormone-related index, cell apoptosis and autophagy during the out-of-season spawning of largemouth bass, and (b) to investigate the protective effect of the antioxidant resveratrol on this fish during out-of-season spawning from May through August. The study was divided into two groups (three replicates per group, 2000 fish per replicate): control group (Control) (exposure to water temperature of 12-17 °C) and resveratrol supplementation group (Res) (exposure to water temperature of 12-17 °C and fed with 200 mg/kg resveratrol). The results show that: (1) The serum hormones LH and E2 increased first and then remained unchanged, and the ovarian section showed that the ovary remained in stage IV. (2) In the process of off-season reproduction, a large number of follicles experienced follicular atresia, accompanied by endoplasmic reticulum expansion, nuclear chromatin condensation and mitochondrial swelling, which was relieved after feeding resveratrol. (3) Resveratrol decreased the ovarian ROS content and improved the activities of CAT and other antioxidant enzymes in the ovary and liver to some extent. (4) Resveratrol reduced the level of pro-apoptotic (Bax, Caspase3, Caspase8, Caspase9) and autophagy-related components (LC3-B, Beclin-1) while increasing the transcription level of anti-apoptotic (Bcl-2) factors. These findings suggest that resveratrol alleviates some adverse effects of largemouth bass during out-of-season spawning to some extent and provide a model for efficient and high-quality out-of-season spawning.

Exposure to Intermittent Environmental Hypoxia Promotes Vascular Remodeling through Angiogenesis in the Liver of Largemouth Bass (Micropterus salmoides).

In this study, we explored the effects of 4 weeks of intermittent hypoxic exposure (IHE) on liver angiogenesis and related regulatory mechanisms in largemouth bass (Micropterus salmoides). The results indicated that the O2 tension for loss of equilibrium (LOE) decreased from 1.17 to 0.66 mg/L after 4 weeks of IHE. Meanwhile, the red blood cell (RBC) and hemoglobin concentrations significantly increased during IHE. Our investigation also found that the observed increase in angiogenesis was correlated with a high expression of related regulators, such as Jagged, phosphoinositide-3-kinase (PI3K), and mitogen-activated protein kinase (MAPK). After 4 weeks of IHE, the overexpression of factors related to angiogenesis processes mediated by HIF-independent pathways (such as nuclear factor kappa-B (NF-κB), NADPH oxidase 1 (NOX1), and interleukin 8 (IL8)) was correlated with the accumulation of lactic acid (LA) in the liver. The addition of cabozantinib, a specific inhibitor of VEGFR2, blocked the phosphorylation of VEGFR2 and downregulated the expression of downstream angiogenesis regulators in largemouth bass hepatocytes exposed to hypoxia for 4 h. These results suggested that IHE promoted liver vascular remodeling by the regulation of angiogenesis factors, presenting a potential mechanism for the improvement of hypoxia tolerance in largemouth bass.

Comprehensive analysis of full-length transcriptomes of Schizothorax prenanti by single-molecule long-read sequencing.

Schizothorax prenanti (S. prenanti) is one of the most important aquaculture species in the southwest of China. However, information of the full-length transcripts in S. prenanti remains unknown. In this study, single-molecule real-time (SMRT) sequencing was performed to generate full-length transcriptomes of S.prenanti. In total, 23.26 Gb of clean reads were generated. A total of 312,587 circular consensus sequences (CCS) were obtained with average lengths of 2634 bp and 84.16% (270,662) of CCS were full-length non-chimeric reads. After being corrected with Illumina library sequencing, 18,005 contigs were obtained, with 17,797 (98.81%) successfully annotated in eight public databases, including 15,839 complete open reading frames (ORFs) with an average length of 1330 bp. Furthermore, a total of 4152 alternative splicing (AS) events and 250 long non-coding RNA (lncRNA) transcripts were detected. Additionally, a total of 1129 putative transcription factors (TFs) members from 56 TF families and 11,660 simple sequence repeats (SSRs) were identified. This study provided a valuable resource of full-length transcripts for further research on S. prenanti.

Yinchenhao Decoction ameliorates the high-carbohydrate diet induced suppression of immune response in largemouth bass (Micropterus salmoides).

Yinchenhao Decoction (YD), a Chinese herbal medicine, has been traditionally used for treatment of metabolic liver diseases. A 10-week feeding trail was carried out to examine the effects of YD supplementation in a high carbohydrate diet (HCD) on liver histopathology, immune response, disease resistance, and expression of genes associated with endoplasmic reticulum stress, autophagy, apoptosis, necroptosis and inflammation in juvenile largemouth. A diet containing 9% carbohydrate was used as a low carbohydrate diet (LCD), and a HCD was formulated to contain 18% carbohydrate and supplemented with 0, 0.5, 1, 2 or 4% YD (HCD, HCD+0.5YD, HCD+1YD, HCD+2YD and HCD+4YD). Triplicate groups of fish (5.6 ± 0.2 g) were feed the test diets to visual satiety for 10 weeks. The highest survival rate after Nocardia seriolae challenge was recorded for the HCD+4YD group. YD application led to reduced ACP, AKP, AST and ALT activities. HCD-induced cells swelling, ruptured cell membrane, migrated nuclei and increasing inflammatory cells in hepatocytes were mitigated by YD addition. Moreover, YD decreased the expressions of pro-inflammation genes (TNF-α, IL-1ß, IL-8, hepcidin1, NF-κB, COX2, CD80 and CD83) and increased the mRNA levels of anti-inflammation genes (IL-10 and IKBα). The mode of liver cell death was preferably changed to programed apoptosis rather than uncontrolled necroptosis by application of YD in HCD. Furthermore, the expression of UPR genes (IRE1, Eif2α, ATF6, XBP1 and GRP78/Bip) and autophagy genes (LC3-2, BNIP3 and P62) was increased by YD supplementation. In summary, our results demonstrated that YD addition in HCD enhances UPR, autophagy and programed apoptosis maintaining the homeostasis, and decreases uncontrolled necroptosis and inflammation, ultimately leading to improved immune response in largemouth bass.

Acute hypoxia promotes the liver angiogenesis of largemouth bass (Micropterus salmoides) by HIF - Dependent pathway.

A 24-h hypoxia exposure experiment was conducted to determine how hypoxia exposure induce liver angiogenesis in largemouth bass. Nitrogen (N2) was pumped into water to exclude dissolved oxygen into 1.2 ± 0.2 mg/L, and liver tissues were sampled during hypoxia exposure of 0 h, 4 h, 8 h, 12 h, 24 h and re-oxygenation for 12 h. Firstly, the results showed that hypoxia exposure promoted the angiogenesis occurrence by immunohistochemical analysis of vascular endothelial growth factor receptor 2 (VEGFR2). Secondly, the concentration of vasodilation factor increased and it's activity was elevated during 8 h exposure, such as nitric oxide (NO) and nitric oxide synthase (NOS) (p < 0.05). Thirdly, hypoxia exposure promoted angiogenesis through up-regulation the expression of matrix metalloproteinase 2 (MMP-2), jagged, protein kinase B (AKT), phosphoinositide-3-kinase (PI3K), mitogen-activated protein kinase (MAPK) at 4 h; contrarily, the expression of inhibiting angiogenesis genes presented up-regulated at 8 h (p < 0.05), such as matrix metalloproteinase inhibitor-2 (TIMP-2), matrix metalloproteinase inhibitor-3 (TIMP-3). Finally, the genes and proteins that regulate angiogenesis presented obvious chronological order. Parts of them promoted the budding and extension of blood vessels were up-regulated during 4 h-8 h (p < 0.05), such as vascular endothelial growth factor a (VEGFA), VEGFR2, monocarboxylic acid transporter 1 (MCT1), CD147, prolyl hydroxylase (PHD), nuclear factor kappa-B (NF-κB); other part of them promoted blood vessel maturation were highly expressed during 12 h-24 h (p < 0.05), such as angiogenin-1 (Ang-1) and angiogenin-2 (Ang-2). In short, acute hypoxia can promote the liver angiogenesis of largemouth bass by HIF - dependent pathway.

Metabolic response provides insights into the mechanism of adaption to hypoxia in largemouth bass (Micropterus salmoides) under intermittent hypoxic conditions.

In metabolism, molecular oxygen is a necessary substrate. Oxygen imbalances are linked to a variety of circumstances in the organism's homeostasis. Recently, the positive effects of hypoxia treatment in improving exercise ability and hypoxia tolerance have become a research focus. We explored the effects of intermittent hypoxia exposure (IHE, for one hour or three hours per day) on the hypoxia tolerance of largemouth bass in this study. The results showed that (1) IHE significantly reduced the LOEcrit (the critical O2 tension for loss of equilibrium) value of largemouth bass, indicating that its hypoxia tolerance was enhanced. (2) The level of oxidative stress in the liver decreased in the HH3 group (exposed to a hypoxic condition for 3 h per day) compared to HH1 group (exposed to a hypoxic condition for 1 h per day). (3) IHE reduced the content of lactic acid and enhanced the process of gluconeogenesis in the liver. (4) Importantly, lipid mobilization and fatty acid oxidation in the liver of largemouth bass were significantly enhanced during IHE. In short, the results of this study indicate that IHE can improve hypoxia tolerance by regulating the energy metabolism of largemouth bass.

Transcriptome Reveals the Effects of Early Weaning on Lipid Metabolism and Liver Health of Yangtze Sturgeon (Acipenser dabryanus).

The Yangtze sturgeon (Acipenser dabryanus) has recently been declared extinct in the wild, and artificial breeding is the only means to protect its germplasm resources, but it has difficulty in weaning (from live prey to artificial food). In this study, we first performed a histological observation, enzyme-activity determination, and transcriptome sequencing on the livers of juvenile Yangtze sturgeons, and we then cloned five critical genes of lipid metabolism according to the transcriptome-sequencing results. We designed a weaning experiment to analyze their expression levels during weaning. The results showed that the density of hepatocytes and the transaminase activity of the juveniles failed to wean. The differentially expressed genes were enriched significantly in the pathways involving steroid synthesis, amino acid metabolism, and pancreatic secretion. It was found that the mRNA level of the fatty acid-synthesis gene decreased, and the mRNA level of the lipolysis gene increased significantly during weaning. The results of this research indicated that weaning could affect the liver health of Yangtze sturgeon, and it could affect the liver lipid metabolism by inhibiting fatty acid synthesis and promoting lipolysis. This study enhances our understanding of the impact of weaning on the lipid metabolism in fish.

Molecular and functional characterization of the retinol-binding protein 4 (RBP4) in hepatocytes of Schizothorax prenanti in response to palmitic acid.

Retinol-binding protein 4 (RBP4) protein is a kind of adipokines synthesized and secreted by the liver, which has been verified to play important roles in liver metabolism and energy homeostasis. However, the effects of RBP4 on hepatic lipid accumulation are still elusive in fish. In the present study, we cloned and characterized the RBP4 gene in Schizothorax prenanti (S. prenanti). RBP4 gene was specifically expressed in the liver and abdominal adipose tissue. Palmitic acid (PA; 400 µM) can significantly increase lipid deposition in primary hepatocytes after 12 h of treatment. Furthermore, RBP4 knockdown can relieve the excessive lipid deposition and endoplasmic reticulum stress in the hepatocytes caused by PA. The inhibition of RBP4 abolished the ability of PA to induce the expression of genes involved in lipogenesis and endoplasmic reticulum stress. These results demonstrate that RBP4 inhibition attenuated PA-induced lipid deposition and endoplasmic reticulum stress in hepatocytes of S. prenanti. This study could contribute to improve the understanding of RBP4 functions in the PA-induced lipid deposition in hepatocytes of fish.

Enhance energy supply of largemouth bass (Micropterus salmoides) in gills during acute hypoxia exposure.

Gills are the location of gas exchange and also the first target organ of fish response for environmental stress. As a multifunctional organ, its energy supply, when faced with insufficient dissolved oxygen in the water, remains unclear. In this study, largemouth bass was subjected to hypoxia stress (1.2 mg/L) for 24 h and 12 h reoxygenation (R12) to evaluate energy supply strategy of gills. Under hypoxia exposure, the respiratory rate of largemouth bass increased by an average of 20 breaths per minute. A total of 2026, 1744, 1003, 579, 485, and 265 differentially expressed genes (DGEs) were identified at 0 h, 4 h, 8 h, 12 h, 24 h, and R12h in gills after hypoxia exposure. KEGG functional analysis of DEGs revealed that the glycolysis/gluconeogenesis pathway was enriched across all the sampling points (0, 4, 8, 12, 24 h, R12). The gene expression and enzyme activity of three rate-limiting enzymes (hexokinase, phosphofructokinase-6, pyruvate kinase) in glycolysis pathway were significantly increased. Increased levels of glycolysis products pyruvate and lactic acid, as well as the number of mitochondria (1.8-fold), suggesting an enhancement of aerobic and anaerobic metabolism of glucose in gills. These results suggest that the gill of largemouth bass enhanced the energy supply during acute exposure to hypoxia stress.

MiRNA-seq analysis of spleen and head kidney tissue from aquacultured largemouth bass (Micropterus salmoides) in response to Aeromonas hydrophila infection.

Recently, the same fish diseases, which have been found in pond farming, have been found in the newly tested largemouth bass (Micropterus salmoides) system. Bacterial septicemia caused by Aeromonas hydrophila occurs frequently in largemouth bass culture leading to significant economic losses. To investigate the role miRNA in the largemouth bass disease resistance, twelve (2 tissues (spleen and head kidney) × 2 experimental groups (infected and control) × three biological replicates) small RNA libraries were constructed and sequenced with miRNA-seq. A total of 26 differentially expressed miRNAs, 8 upregulated and 18 downregulated, were identified in the spleen, and 19 differentially expressed miRNAs, 9 upregulated and 10 downregulated, were identified in head kidney (fold change ≥ 2 or ≤ 0.5 and P ≤ 0.05). The differentially expressed miRNAs with the largest fold change were selected for target gene prediction using GO and KEGG analysis. Six miRNAs in the spleen and 5 miRNAs in the head kidney were selected. Analysis showed that, of all the immune and metabolic pathways, the FoxO signaling pathway was enriched in both the spleen and head kidney groups. Common target genes of the pathway included AMP-activated catalytic subunit alpha 1 (prkaa1), phosphatidylinositol 3-kinase (pik3r3b), serine/threonine-protein kinase (plk2), and forkhead box protein G1 (foxg1a). MiRNAs (such as miR-126-5P, miR-126-3P) are involved in immune response and cell cycle functions as they regulate targeted genes in the FoxO pathway. These results will enhance our understanding of the molecular mechanisms underlying immune responses to bacterial septicemia and facilitate molecular-assisted selection of resistant strains of largemouth bass.

High carbohydrate diet induced endoplasmic reticulum stress and oxidative stress, promoted inflammation and apoptosis, impaired intestinal barrier of juvenile largemouth bass (Micropterus salmoides).

This study assessed the effects of feed carbohydrate content on intestinal physical barrier and immunity in juvenile largemouth bass (Micropterus salmoides). Triplicate groups of juvenile fish (4.1 ± 0.2 g) were fed low (LCD, 7%), medium (MCD, 12%) and high (HCD, 17%) carbohydrate diets for eight weeks. Gut histology revealed the slight infiltration of inflammatory cells and moderate loss of mucous membrane layer in HCD group. Expression of ZO1, occluding, and claudin7 genes and epidermal growth factor receptor (EGFR) gene were significantly decreased in HCD group indicating the impairment of tight junction and epithelial cell regeneration. The results showed the significant (P < 0.05) reduction of antioxidant capacity in HCD group compared to LCD. Furthermore, expression of intestinal ERS-related genes such as IRE1, Eif2α, GRP78, CHOPα and CHOPß in HCD group was significantly higher than the LCD group. In addition, HCD induced the up-regulated expression of inflammatory (IL-8, IL-1ß, TNFα and COX2) and apoptosis (TRAF2, bax, casepase3, caspase8 and casepase9) related genes in fish intestine. The data generated in this study clearly demonstrated that HCD induced ERS and oxidative stress, which promoted intestinal inflammation and apoptosis in juvenile largemouth bass.

Interactive effect of thermal and hypoxia on largemouth bass (Micropterus salmoides) gill and liver: Aggravation of oxidative stress, inhibition of immunity and promotion of cell apoptosis.

High temperatures and low oxygen in aquatic environments, such as intensive aquaculture or in natural watersheds, inevitably cause stress in fish. Fish are exposed to high temperatures during the summer, which exacerbates hypoxia. Hypoxia (1.2 ± 0.2 mg/L) under 20 °C (20 HG) and 26 °C (26 HG) was simulated to induce stress in largemouth bass (Micropterus salmoides). Related enzymes and genes involved in antioxidant, immune, and apoptotic responses were selected to explore the interactive effects of temperature and hypoxia on largemouth bass. The results showed that malondialdehyde (MDA) levels in plasma, gill, and liver increased in the 26 HG (p < 0.05). Liver superoxide dismutase (SOD) activity increased in the 26 HG. Peak SOD (SOD1, SOD2, SOD3a, and SOD3b), CAT, and GSH-Px mRNA levels in the gill and liver were observed at 12-24 h of stress. The levels of gill and liver total antioxidant capacity, catalase (CAT), glutathione peroxidase (GSH-Px) activities and other enzyme activities and genes in the 26 HG were higher than those in the 20 HG (p < 0.05). The gill and liver acid phosphatase and alkaline phosphatase activities increased with time in the 26 HG (p < 0.05), while gill and liver lysozyme activities in the 26 HG were lower than those in the 20 HG (p < 0.05). Tumor necrosis factor-α mRNA level was upregulated in the gill and downregulated in the liver at 24 h in the 26 HG. Interleukin (IL)-1ß and IL-8 mRNA levels were upregulated in the gill and liver in the 26 HG at 24 h, whereas IL-15 mRNA level was downregulated in the 26 HG at 12 h. Transforming growth factor-ß1 mRNA level was upregulated in the gill in the 20 HG at 24 h, but downregulated in gill and liver in the 26 HG at 24 h. Similarly, IL-10, Hepcidin-1, and Hepcidin-2 showed lower expression levels in the 26 HG. Gill and liver caspase-3 activities were higher in the 26 HG (p < 0.05), and gill caspase-3 activity was higher than that in the liver. The mRNA levels of proapoptotic genes (caspase-3, caspase-8, and caspase-9) were higher in the 26 HG. The present study demonstrates the interactive effects of temperature and hypoxia on stress in largemouth bass gill and liver. These results will be helpful to understand the mechanisms of stress induced by temperature and hypoxia in fish and provide a theoretical basis for aquaculture management.

Rice-shrimp culture: a better intestinal microbiota, immune enzymatic activities, and muscle relish of crayfish (Procambarus clarkii) in Sichuan Province.

Intestinal microbiota play an important role in the intestinal immunity and nutrient absorption, even muscle nutritional components, and the composition and function were affected by environment. In this study, the intestinal microbiota and immune enzyme, nutritional flavor of muscle of crayfish in rice field, and pond cultivation model were compared in summer and autumn. The results of Shannon diversity and Chao 1 index of intestinal microbiota based on 16S sequencing analysis showed that the diversity and abundance in autumn were higher than in summer. And the diversity and abundance of intestinal microbiota of different model in the same season were different. Four dominant phyla (relative abundance > 5% at least in one sample) of the intestinal microbiota were Bacteroidetes, Firmicutes, Proteobacteria, and Tenericutes. From summer to autumn, the intestinal immune enzyme activity of crayfish in both models showed a decreasing trend. In summer, the activity of catalase and alkaline phosphatase of crayfish cultured in the pond was significantly higher than that in rice field (P < 0.05). In autumn, the activity of catalase and lysozyme of crayfish cultured in rice field was significantly higher than that in pond (P < 0.05). The contents of umami and sweetish amino acids of muscle were higher in rice field than in pond, and the percentage of glutamic acid and alanine was significantly higher in rice field than in pond (P < 0.05). Thus, rice field model can make crayfish a more stable intestinal environment and a better intestinal immune enzyme activity and muscular flavor. Key points • The intestinal microbiota of crayfish in rice field had tended to stabilize from summer to autumn. • The crayfish had better nutrient absorption and stronger immune abilities in the rice field. • The crayfish cultured in rice field had higher overall umami concentration than in pond.

Characterization, tissue distribution of resistin gene and the effect of fasting and refeeding on resistin mRNA expression in Siberian sturgeon (Acipenser baerii).

Resistin as an adipokine identified from rodents in 2001 is involved in many biological processes. However, little is known about this gene in fish. We cloned Siberian sturgeon (Acipenser baerii) resistin cDNA of 795 base pairs, encoding 107 amino acids, which showed 38-40% identity to human and rodents. Real-time quantitative PCR showed that the resistin gene was widely distributed in tissues of Siberian sturgeon, with the highest expression in liver. After fasting for 1, 3, 6 and 10 days, the expression of the resistin gene in the liver of Siberian sturgeon decreased significantly, and after refeeding on the 10 days of fasting the resistin mRNA expression increased rapidly, suggesting that resistin may play an important role in liver in response to starvation. Taken together, these results suggest that resistin may be involved in the regulation of energy homeostasis in liver.

Analysis of miRNA-seq in the liver of common carp (Cyprinus carpio L.) in response to different environmental temperatures.

Water temperature affects the survival, growth, immunity, reproduction, and productivity of farmed fish. The temperature beyond suitable range will disrupt the normal physiological activity. Common carp (Cyprinus carpio L.) is a representative eurythermic fish; they are able to sense and respond to changes in water temperature by adjusting their physiology. To investigate the miRNAs in common carp at different temperatures, nine liver small-RNA libraries (5 °C, 17 °C, and 30 °C, each group have three biological repetitions) were constructed and sequenced using high-throughput sequencing. A total of 110 known miRNAs were identified. Twenty-nine known miRNAs were differentially expressed compared with in control group. GO and KEGG analysis indicated that the miRNAs may play important roles in metabolism and environment information processing. Specifically, we considered the insulin-signaling and glycerophospholipid metabolism pathway, and the results show that in 30 °C, miR-301a, miR-203b-5p, and miR-210-3p were upregulated; their target genes which are the mechanistic targets of the rapamycin kinase (mtor) gene and the protein kinase AMP-activated catalytic subunit alpha 1 (prkaa1) gene in the insulin-signaling pathway were downregulated. And miR-9-5p, miR-27d, miR-92b-3p, and miR-155 were upregulated; their target genes, 1-acylglycerol-3-phosphate O-acyltransferase 3 (agpat3), CDP-diacylglycerol-inositol 3-phosphatidyltransferase (cdipt), glycerol-3-phosphate acyltransferase mitochondrial (gpam), and phosphatidylglycerophosphate synthase 1 (pgs1), in glycerophospholipid metabolism pathway were downregulated. But in 5 °C, the situation was opposite. These findings suggest that significant changes occur in energy metabolism and metabolic processes with components of the cell membrane in different temperatures, which significantly advance our understanding of the regulatory mechanisms underlying the physiological change of temperature stress-induced in liver, specifically with regard to miRNAs. These data provide a foundation for further studies of the role of miRNAs in environmental adaptation in fish.

Influence of long-term temperature stress on respiration frequency, Na+/K+-ATPase activity, and lipid metabolism in common carp (Cyprinus carpio).

Temperature is a key factor that influences fish metabolism. Therefore, it is important to understand how temperature variation affects lipid metabolism and energy consumption in fish. We determined respiration frequency, Na+/K+-ATPase activity, and lipid metabolism in common carp (Cyprinus carpio) after 18 days of temperature stress (5 °C and 30 °C, with 17 °C as the control). We found that respiratory frequency was positively correlated with water temperature (p < 0.01), whereas red blood cell count and mean corpuscular hemoglobin content and concentration were negatively correlated with temperature. In liver and heart, triglyceride (TG) levels were lower in both stress groups (5 °C and 30 °C) than in the control (17 °C) (p < 0.01), and highest in muscle from the high temperature stress group (p < 0.01). The non-esterified fatty acid concentration was negatively correlated with TG levels. In brain, lipoprotein lipase (LPL) activity decreased with increasing temperature and hepatic lipase (HL) activity was lower in both stress groups than in the control (p < 0.01). In muscle, the activity of LPL and HL was lowest in the high temperature stress group, resulting in a significant increase in TG levels. Na+/K+-ATPase activity in heart was lower in both stress groups than in the control (p < 0.01). The expression of LPL mRNA in heart increased with increasing temperature (p < 0.01), whereas LPL mRNA expression in brain and liver increased in both stress groups (p < 0.01). Our results show that temperature can significantly affect lipid metabolism in common carp, and that different tissues respond differently to changes in temperature.

GSK3ß inhibition suppresses the hepatic lipid accumulation in Schizothorax prenanti.

Glycogen synthase kinase-3ß (GSK3ß) is a serine/threonine kinase involved in the regulation of embryonic development, glycogen metabolism, protein synthesis, mitosis, and apoptosis. To understand the role of GSK3ß in hepatic lipid accumulation of Schizothorax prenanti, we used lithium chloride (LiCl), a GSK3ß inhibitor, to inhibit the expression and activity of GSK3ß. LiCl increased levels of phosphorylation of GSK3ß (Ser9) and decreased the protein level of GSK3ß. Plasma TG, TC, and LDL-C levels were greatly decreased after LiCl treatment. Additionally, GSK3ß inhibition significantly reduced the levels of hepatic triglyceride (TG) and decreased the expression of lipogenesis-related genes in liver. Interestingly, LiCl decreased levels of phosphorylation of STAT3 (Tyr705), and then inhibited the activity of STAT3. These results indicate that in vivo LiCl treatment, which inhibited GSK3ß activity, effectively decreased hepatic lipid accumulation through STAT3 in Schizothorax prenanti.

Effect of starvation and refeeding on oxidative stress and antioxidant defenses in Yangtze sturgeon (Acipenser dabryanus).

The present research aimed to evaluate the effects of long-term fasting and refeeding on the growth and antioxidant defenses in the liver and serum in Yangtze sturgeon (Acipenser dabryanus). The results showed that body mass and hepatosomatic index significantly decreased with long-term fasting, but they could be recovered after 4 weeks refeeding. Compared with controls, the antioxidant defense parameters of starvation indicated that the malondialdehyde (MDA) levels increased significantly in both tissues; the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) increased obviously in serum and liver, respectively (p < 0.05). However, the activities of catalase (CAT) always decreased in two tissues including liver and serum during the whole starvation, as was the SOD in the liver (p < 0.05). Interestingly, the T-AOC levels of Yangtze sturgeon presented higher at early stage of starvation and dropped down at the end of starvation (p < 0.05). However, all of the antioxidant index above returned to origin level after 4 weeks refeeding. In conclusion, the present study indicated that long-time fasting induced oxidative stress in Yangtze sturgeon and it may easily adjust their physiological status under situations characterized by a long-term starvation and refeeding.

Differences in the digestive enzyme activity, intestinal mucosa and microbial community in loach cultivated in two separate environments.

BACKGROUND: Fish culture in rice paddies can contribute to increasing yields of rice and surplus fish products. Environmental impacts and food-safety issues have become important topics in aquaculture, and organic foods currently were paid attention by researchers and industry practitioners. But the mechanism of differences in quality of Loach (Paramisgurnus dabryanus) reared in rice fields and ponds remains largely uncharacterized. In this study,digestive enzyme activity, intestinal mucosa cells and the gut microbial community of loach were determined under the two separate cultivation modes. RESULTS: The levels of intestinal digestive enzyme activity of fish reared in the paddy-cultivated mode (PACM) were higher (P < 0.05) than those in the pond-cultivated mode (POCM). It was extremely significant (P < 0.01) for the activity of lipase in the liver, foregut and midgut, and for the activities of amylase and trypsin in the hindgut. Acid mucous cells in the loach foregut in PACM were fewer than in POCM (P < 0.01). In summer, the abundance of the Firmicutes, Lactobacillus spp., Aeromonas hydrophila, Enterobacteriaceae and Streptococcus spp. in loach intestinal mucosa in PACM was higher than in POCM. In fall, the abundance of total bacteria, the Bacteroidetes, Bifidobacterium and Enterobacteriaceae in the intestinal mucosa in PACM was likewise higher than in POCM. These differences were significant (P < 0.05 or P < 0.01) between loach in the two separate culture modes for all microorganisms except for A. hydrophila and Streptococcus spp. In addition, quantitative PCR assays showed that some microorganisms presented consistently similar abundances in the gut as in the culture water. CONCLUSIONS: These results showed some enzymatic activities involved in digestion in liver and intestine of loach in PACM were higher than those in POCM, as using digestive enzyme analysis and histological observation of intestinal sections. These findings suggest most of the microorganisms examined in the gut mucosa of loach in the two culture modes significantly differed in abundance between summer and fall. However, some pathogenic bacteria in the gut, particularly A. hydrophila, presented lower abundance in PACM in fall, yet did not differ in abundance between loach in the two cultivation modes.

Histopathological effects of Pedunsaponin A on Pomacea canaliculata.

Pedunsaponin A, a novel molluscicidal compound isolated from Pueraria peduncularis, exhibits strong toxicity against Pomacea canaliculata. To determine the mechanisms of Pedunsaponin A toxicity, its effects on the organs and hemocytes of P. canaliculata were examined in this study. The results showed that Pedunsaponin A had significant toxic effects on different organs of the snail, including the lungs, gills, mantle, siphon tube, ventricle, pericardial cavity, hepatopancreas, kidneys, and the major symptom of this toxicity was the loss of cilia in the lungs and gills. Additionally, in further studies on the effects of Pedunsaponin A treatment, we found that the hemocyte count was changed and hemocyte morphology was damaged, which was primarily reflected by cytoplasm leakage, nuclei deformation, and significant reductions in the number of ribosomes and granulocyte mitochondria. Based on these results and considering that blood vessels are distributed in the lungs and gills, we hypothesized that Pedunsaponin A would first destroy the cilia, which disrupt physiological activities such as respiration, excretion and feeding, and then enter the hemolymph through blood vessels, disrupt the normal function of the hemocytes and destroy the snail immune system, eventually resulting in the death of the snail.