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1.
The immune response and morphological changes in the gills of rainbow trout fry after immersion in hydrogen peroxide (H2O2), Flavobacterium psychrophilum or combined exposure were examined. The gills were sampled 4, 48, 125 and 192 h after exposure, and the regulation of expression of the following genes was investigated using qPCR: IgT, IgM, CD8, CD4, MHC I, MHC II, IL-4/13A, TcR-β, IL-10, IL-1β, IL-17, SAA and FoxP3. Bacteria were not observed in haematoxylin-and-eosin-stained gill tissue, but the presence of F. psychrophilum 16S rRNA was detected using qPCR. The 16S rRNA levels were correlated with gene expression. Although pretreatment with H2O2 before immersion in F. psychrophilum did not significantly alter the amount of bacteria found in the gill, the immune response was influenced: exposure to F. psychrophilum resulted in a negative correlation with expression of IL-17c1, MHC I and MHC II, while pretreatment with H2O2 resulted in a positive correlation with IL-4/13A and IgM. Exposure to either H2O2 or F. psychrophilum influenced the regulation of gene expression and damaged tissue. Exposure to both combined altered the immune response to infection and postponed healing of gill tissue.  相似文献   

2.
The toxicity of hydrogen peroxide (H2O2) to fingerling (80-160 mm total length) walleye, Stizostedion vitreum, was evaluated. Walleye were exposed in a 1 hour static bath to 50, 75, 100, 200, 225, 250, 400, and 600 ppm (µL/L) active ingredient (H2O2) in five experiments. Three to six replicates of each concentration were used in each experiment, with 25 fish per replicate. After resuming flow, concentrations of H2O2 in the tanks declined by 23.8% every hour, reaching 0 ppm (µL/L) about 3.5 hours after treatment. Mortalities, as well as dissolved oxygen, temperature, and H2O2 concentrations were monitored at 1, 12, 24, 48, 72, and 96 hours. Dissolved oxygen concentrations increased 0.25-1.16 ppm (µL/L) 1 hour after the addition of H2O2, because of the dissociation of H2O2 (2H2O2→2H2O + O2). The LC50 following a single 1-hour exposure ranged from 145.1 ppm (µL/L) at 12 hours to 142.8 ppm (µL/L) at 96 hours, which indicates that the toxic effect is mainly from initial contact. The toxicity threshold for walleye is about 50 ppm (µL/L) (1.3% mortality at 96 hours following 1 hour exposure). Other studies indicate that rainbow trout, Oncorhynchus mykiss, tolerate up to 250 ppm (µL/L) for 1 hour, but fingerling walleye should not be exposed to more than 50 ppm (µL/L) for the same exposure interval.  相似文献   

3.
The immune response in rainbow trout fry against Flavobacterium psychrophilum was elucidated using an immersion‐based challenge with or without prior exposure to hydrogen peroxide (H2O2). Samples were taken from the head kidney 4, 48, 125 and 192 h after immersion, and the regulation of several genes was examined. Bacterial load was assessed based on the presence of 16S rRNA and correlated with gene expression, and the levels of specific antibodies in the blood were measured 50 days post‐infection. Separately, both H2O2 and F. psychrophilum influenced gene expression, and pre‐treatment with H2O2 influenced the response to infection with F. psychrophilum. Pre‐treatment with H2O2 also affected correlation between gene regulation and pathogen load for several genes. A delay in antibody production in H2O2‐treated fish in the early phase of infection was indicated, but H2O2 exposure did not affect antibody levels 50 days post‐infection. An increasing amount of F. psychrophilum 16S rRNA was found in the head kidneys of infected fish pre‐treated with H2O2 relative to the F. psychrophilum group. The results show that a single pre‐treatment with H2O2 impairs the response against F. psychrophilum and may intensify infection.  相似文献   

4.
Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease and rainbow trout syndrome in freshwater salmonid fish worldwide, generating injuries and high mortality rates. Despite several studies on this bacterium, the infection mechanism remains unknown due to limitations in the employed animal models. In this work, we propose using zebrafish (Danio rerio) as a model for studying bacterial pathogenicity. To substantiate this proposal, zebrafish infection by F. psychrophilum strain JIP 02/86 was characterized. Zebrafish larvae were infected using the bath method, and morphological changes and innate immune system activation were monitored using transgenic fish. Salmonid-like infection phenotypes were observed in 4.74% of treated larvae, as manifested by fin, muscle and caudal peduncle damage. Symptomatic and dead larvae accounted for 1.35% of all challenged larvae. Interestingly, infected larvae with no infection phenotypes showed stronger innate immune system activation than specimens with phenotypes. A failure of function assay for myeloid factor pu.1 resulted in more infected larvae (up to 43.5%), suggesting that low infection rates by F. psychrophilum would be due to the protective actions of the innate immune system against this bacterium in zebrafish larvae. Our results support the use of zebrafish as an infection model for studying F. psychrophilum. Furthermore, the percentage of infected fish can be modulated by disturbing, to varying extents, the differentiation of myeloid cells. Using this evidence as a starting point, different aspects of the infection mechanism of F. psychrophilum could be studied in vivo.  相似文献   

5.
This study was conducted to investigate the protective effect of L‐carnitine (LC) against H2O2‐induced oxidative stress in the fathead minnow muscle cell line (FHM). The FHM cells were stimulated with 1 mM H2O2 for 1 h after LC pre‐treatment, and the cell viability and the activity and mRNA relative expression of antioxidant enzyme were measured to assess the antioxidant properties of LC. The results showed that the toxic effect of H2O2 on the viability of FHM cells was both dose‐ and time‐dependent. Furthermore, the viability of the 0.01–1 LC mM groups was significantly higher than those of the 1 mM H2O2 group. L‐carnitine protected the cells from H2O2‐induced oxidative damage, which was demonstrated by a significant reduction in the malondialdehyde and reactive oxygen species levels and increases in the intracellular total glutathione levels and the activities of total superoxide dismutase, catalase, glutathione peroxidase (GPx) and gamma‐glutamyl‐cysteine synthetase (γ‐GCS) in FHM cells pre‐treated with LC for 6 h compared with the 1 mM H2O2 group. In addition, the mRNA relative expression levels of the γ‐GCS catalytic subunit and nuclear factor nuclear factor erythroid 2‐related factor 2 were significantly higher than those of the 1 mM H2O2 group. It could be concluded that LC exerts a beneficial antioxidant effect against oxidative stress induced by H2O2 in FHM cells and that the appropriate treatment is 0.1–1 mM for 6 h in this study.  相似文献   

6.
A fish meal supply shortage is limiting aquaculture development. Currently, plant‐based proteins, such as soya bean meal, are being used as an alternative protein source, despite that such a diet can adversely affect fish, such as by inducing an inflammatory response. A possible solution is to include dietary additives in farm diets to counteract negative effects. One such solution originates from pine bark extracts, which present bioactive properties. In this study, the antioxidant and anti‐inflammatory properties of Pinus radiata bark extracts were evaluated for the first time in a salmonid cell line. This extract chemically demonstrated antioxidant activity through 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH = 58.4 ± 1.1%) and ferric ion reducing antioxidant power (FRAP = 575 ± 17 mgEqFe(II)·g extract?1) assays. Additionally, the extract showed high flavonoid and phenolic compound contents. Up to 100 mg mL?1, the P. radiata extract showed no cytotoxicity in the CHSE‐214 salmonid embryo cell line. Moreover, the antioxidant activity of the extract (50 μg mL?1) was evaluated by a dichlorofluorescein (DCFH) assay in the SHK‐1 salmon cell line challenged with an oxidant stimulus (H2O2), showing 58.9% activity. The extract also protected DNA from oxidative damage, as observed through a comet assay. When assessing anti‐inflammatory properties in an in vitro inflammation model, the extract significantly reduced the relative expression of the pro‐inflammatory cytokines interleukin‐1β (IL‐1β), tumour necrosis factor‐α (TNF‐α) and interleukin‐8 (IL‐8) and of the inducible cyclooxygenase‐2 (COX‐2) enzyme. These results suggest a potential application of P. radiata bark extract in functional foods in aquaculture.  相似文献   

7.
四带小鲃(Puntius tetrazona)是一种具有较高经济价值的小型热带观赏鱼。我们对野生型和长鳍突变型四带小鲃的体长、尾鳍长、尾鳍分节数目和分节长度共4个指标进行了统计分析。结果显示,6月龄野生型(n = 30)与突变型(n = 30)成鱼体长没有显著差异(P = 0.904 2),但两者在尾鳍长、尾鳍分节数目和分节长度3个指标存在显著差异(P < 0.000 1)。同时,我们对不同体长野生型(n = 80)和突变型(n = 80)尾鳍长度、分节数目和分节长度进行了线性回归分析,结果显示,相对于野生型,突变型尾鳍长度增加速率更快(野生型:k = 0.28,突变型:k = 0.62,P < 0.000 1),尾鳍分节数目增加频率基本一致(野生型:k = 0.79,突变型:k = 0.82,P = 0.006),尾鳍分节长度更长(野生型:k = 0.006 8,突变型:k = 0.000 4,P < 0.000 1)。野生型和突变型成鱼尾鳍组织的转录组分析结果显示,转录组从头组装共获得56 271个unigene,野生型组(n=3)和突变型组(n=3)之间共有1 304个差异表达基因,其中971个基因上调,333个基因下调。差异表达基因 KEGG通路显著富集到细胞周期和DNA复制等与细胞增殖相关的信号通路,其中细胞周期蛋白(cyclin A,cyclin B和cyclin E)和细胞周期依赖性激酶(cdk1)等基因显著上调。最后,对ckap2l、ki-67、cdc20、prc1、urgcp和cdks共6个基因的表达水平进行了qRT-PCR分析,其结果与转录组分析基本一致。以上结果表明,长鳍四带小鲃突变基因引起细胞周期相关基因表达量上升,造成尾鳍分节片段的细胞增殖速率加快,尾鳍分节长度增加,呈现为长尾鳍表型。本研究结果为鉴定突变基因和揭示长尾鳍形成的分子机制奠定基础。  相似文献   

8.
Effects of adrenaline on the equilibrium distributions of Na+ , K+ , H+ , Cl , and H2O across the cell membrane of rainbow trout (Salmo gairdneri) erythrocytes were determinedin vitro, as a function of P CO2 (1.76–7.77 torr). CO2-carrying capacity of the blood was also examined. Plasma catecholamine concentrations inunanaesthetized, unrestrained trout were 3.1 nM adrenaline and 1.2 nM noradrenaline. Elevation of the plasma adrenaline concentrationin vitro to 4.6 × 103 nM resulted in net gains of Na+ , Cl and H2O by red cells, a net loss of H+ from red cells, and a pronounced red cell swelling. Adrenaline also reduced the CO2-carrying capacity of trout bloodin vitro. The magnitudes of these effects increased with PCO2 and, thus, were sensitive to blood HCO3 concentrations. The distribution of K+ between red cells and plasma was unaffected by adrenaline. Adrenergic-mediated ion movements and red cell swelling were sensitive to both propranolol and SITS. These results are consistent with the symport NaCl uptake model for adrenergic-mediated swelling of Baroinet al. (1984). The adrenergic response of fish erythrocytes may function to ameliorate the effects of blood acidoses on O2-carrying capacity by maintaining red cell pH in the face of a decrease in plasma pH.  相似文献   

9.
本研究建立了锦鲤(Cyprinus carpio)尾鳍细胞系。染色体数目、核型及DNA含量等实验,发现锦鲤体细胞和锦鲤培养细胞无显著性差异,染色体数目和DNA含量符合比例关系,建立的锦鲤尾鳍细胞系已形成了稳定的遗传性状,命名为KF-H。  相似文献   

10.
This study proposes a new and simple assay that allows rapid assessment of microbial activity in water samples. The assay consists of standardized hydrogen peroxide (H2O2) addition to a water sample and subsequent spectrophotometric determination of H2O2 reduction over time. The H2O2 decomposition rate constant reflects the level of enzymatic activity from planktonic and particle-associated bacteria as well as algae and protozoans. The proof of concept was verified on water samples from recirculating aquaculture systems (RAS), showing that the vast majority of H2O2 decomposition was related to microbial activity. Only 3% of the total H2O2 decomposition was related to abiotic processes when 0.20 μm sterile filtered RAS water was compared with unfiltered RAS water. Planktonic bacteria (size range 0.20–1.6 μm) accounted for 16% of H2O2 decomposition, while bacterial aggregates, particle-associated bacteria and microbiota above 1.6 μm were responsible for the remaining 81%. H2O2 decomposition rate constants were positively correlated to BOD5 (r = 0.893; p < 0.001; n = 18) and to the number of 1–30 μm micro particles (r = 0.909; p < 0.001; n = 72) in RAS water, substantiating the biologically mediated decomposition processes in the water phase. The H2O2 decomposition assay thus represents a new alternative to existing methods that allows rapid (1–2 h) and simple quantification of microbial activity in fresh- and saltwater samples from aquaculture systems. Potential applications of the assay are discussed.  相似文献   

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