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1.
An exopolysaccharide(EPS) was isolated and purified from an Antarctic psychrophilic bacterium B-3,identified as Psychrobacter sp.,and the activation of RAW264.7 cells by B-3 EPS was investigated.The results show that B-3 EPS,over a certain concentration range,promoted cell viability,nitric oxide production,tumor necrosis factor(TNF)a secretion,and phagocytic ability.Furthermore,TAK-242,an inhibitor of the toll-like receptor 4(TLR4) significantly reduced nitric oxide production by these cells after stimulation with B-3 EPS.Moreover,B-3 EPS induced p65 phosphorylation and IκBα degradation in these cells.In conclusion,B-3 EPS might have activated RAW264.7 cells by combining with TLR4 on cell surface and triggering activation of NF-κB signaling pathways,implying that this EPS could activate macrophages and regulate initial immune response.  相似文献   

2.
Recently,the use of natural immunopotentiators,such as plant extracts,is predicted to offer high disease-prevention potential in aquaculture.However,few reports on the immunomodulatory impacts of olive extract(OE)on teleost are available.Therefore,the effects of dietary intake of triterpene-enriched OE on black sea bream(Acanthopagrus schlegelii)were investigated in this study.Our data showed that total blood cell counts,in vivo content of lysozyme,activities of antiprotease and myeloperoxidase,and contents of IL-4 and IL-6 were significantly up-regulated by dietary intake of triterpene-enriched OE.Additionally,the hampered immune response induced by cadmium exposure was significantly mitigated by the administration of OE as were indicated by partially or completely rescued immune-relating parameters.Furthermore,the expressions of immune-related genes encoding NF-κB inhibitor alpha(IkBα),tumor necrosis factorα(TNFα),cyclooxygenase-2(COX-2),and proto-oncogene protein c-fos(FOS)were found to be significantly up-regulated by the dietary intake of OE.In general,the results suggested that the dietary intake of triterpene-enriched OE has immune enhancing effect in black sea bream.Such effect may be realized by 1)increasing the total counts of diverse blood cells;2)activating nonspecific immune biomolecules;and 3)affecting signaling pathways such as IKK and ERK and subsequently inducing IL-4 and IL-6 biosyntheses.  相似文献   

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5.
To investigate the possible use of waste products obtained after processing haddock, the present study prepared haddock bone calcium powder by NaOH and ethanol soaking (alkalinealcohol method) and prepared haddock bone calcium tablets using the powder in combination with appropriate excipients. The biological efficacy of the haddock bone calcium tablets was investigated using Wistar rats as an experiment model. Results show that the optimal parameters for the alkalinealcohol method are: NaOH concentration 1 mol/L, immersion time 30 h; ethanol concentration 60%, immersion time 15 h. A mixture of 2% polyvinylpyrrolidone in ethanol was used as an excipient at a ratio of 1:2 to full-cream milk powder, without the use of a disintegrating agent. This process provided satisfactory tablets in terms of rigidity and taste. Animal studies showed that the haddock bone calcium tablets at a dose of 2 g·kg-1·d-1 or 5g·kg-1·d-1 significantly increased blood calcium and phosphorus levels and bone calcium content in rats. Therefore, these tablets could be used for calcium supplementation and prevent osteoporosis. Although the reasons of high absorption in the rats fed with haddock bone calcium tablets are unclear, it is suggested that there are some factors, such as treatment with method of alkaline-alcohol or the added milk, may play positive roles in increasing absorption ratio.  相似文献   

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7.
A growth experiment was conducted on cobia(Rachycentron canadum,initial weight 108.2 g ± 3.0 g) to investigate the effects of dietary corn gluten meal(CGM) levels on the fish growth,whole body composition and protein metabolism in relation to specific gene expression.Five isonitrogenous(crude protein 45%) and isoenergetic(gross energy 20 kJ g 1) practical diets were formulated by replacing 0%(the control),17.5%,35.0%,52.5%,and 70.0% of fish meal(FM) protein with CGM protein.No significant differences were observed in the survival,feed intake(FI),specific growth rate(SGR),feed efficiency(FE) and protein productive value(PPV) among fish fed diets with 0%,17.5%,35.0%,and 52.5% of CGM protein.However,these indices were significantly lower in fish fed the diet with 70.0% of CGM protein than those in fish fed the control diet(P < 0.05).The whole-body crude protein and lipid contents were significantly lower while the whole-body moisture content was significantly higher in fish fed the diet with 70.0% of CGM protein compared with the control group(P < 0.05).When 70.0% of FM protein was replaced by CGM,plasma total protein and cholesterol contents were significantly lower than those in the control group(P < 0.05).Fish fed the diet with 70.0% of CGM protein had significantly lower hepatic insulin-like growth factor I(IGF-I) expression levels than those in the control group(P < 0.05).However,no significant differences were observed in hepatic target of rapamycin(TOR),dorsal muscle IGF-I and TOR expression levels among dietary treatments.Results of the present study indicated that 52.5% of FM protein could be replaced by CGM in the diets without significant influences on the growth,feed utilization and protein metabolism of juvenile cobia.The present results might be useful for developing cost effective and sustainable cobia dietary formulations.  相似文献   

8.
Low molecular weight peptide (LMWP) was prepared from clam Paphia undulate and its antiaging effect on D-galactose-induced acute aging in rats, aged Kunming mice, ultraviolet-exposed rats, and thermally injured rats was investigated. P. undulate flesh was homogenized and digested using papain under optimal conditions, then subjected to Sephadex G-25 chromatography to isolate the LMWP. Administration of LMWP significantly reversed D-galactose-induced oxidative stress by increasing the activities of glutathione peroxidase (GPx) and catalase (CAT), and by decreasing the level of malondialdehyde (MDA). This process was accompanied by increased collagen synthesis. The LMWP prevented photoaging and promoted dermis recovery and remission of elastic fiber hyperplasia. Furthermore, treatment with the LMWP helped to regenerate elastic fibers and the collagen network, increased superoxide dismutase (SOD) in the serum and significantly decreased MDA. Thermal scald-induced inflammation and edema were also relieved by the LWMP, while wound healing in skin was promoted. These results suggest that the LMWP from P. undulate could serve as a new antiaging substance in cosmetics.  相似文献   

9.
Phycoerythrin (PE) is one of the most important proteins involved in light capturing during photosynthesis in red algae. Its potential biological activities had gained wide concerns. In the present study, tumor cytotoxic and hydroxyl radical assay were preformed to detect the bioactivity of recombinant PE. Recombinant plasmids pGEX-PE and pBGL were transformed into E.coli BL21 to make two recombinant strains BEX (pGEX-PE) and BGL (pBGL). PE expressing in BEX (pGEX-PE) was validated by SDS-PAGE and Western blotting analysis. SDS-PAGE analysis indicated that the PE-GST fusion protein was mostly inclusion bod- ies. Specific expression of PE was confirmed by Western blotting analysis. The recombinant E. coli BEX (pGEX-PE) cells were col- lected and sonicated. The supernatants were reserved for the tumor cytotoxic experiments. The result of tumor cytotoxic assay indi- cated that the supernatants containing PE had the activity of inhibiting the growth of Hela cells and with the increase of protein con- centration, the inhibiting rate increased from 37.31% to 63.26%, which showed significant difference from the control. Hydroxyl radical scavenging effect was tested with supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates treated with sonication and heating. For the sonication samples, the scavenging rates of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were significantly higher than the negative control BL21(pGEX-4T) (P<0.02), and the scavenging rates increased slowly following the increase of the protein content. For the heating samples, except for the 0.2 mg mL-1 BGL (pBGL) products, the scavenging effects of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were stronger than that of negative control BL21(pGEX-4T). However, the effect intensity was not positively correlated with the increase of the protein concentration. Though a partially de- creased hydroxyl radical scavenging activity was led by heating, the biological activity was still retained and conspicuous. This re- search showed that phycoerythrin protein expressing in E. coli has the potential medical and sanitarian value.  相似文献   

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11.
Increasing evidence indicates that transforming growth factor β(TGF-P) signaling pathways play many important roles in the early development of mollusks.However,limited information is known concerning their detailed mechanisms.Here,we describe the identification,cloning and characterization of two Smad genes,the key components of TGF-P signaling pathways,from the Pacific oyster Crassostrea gigas.Sequence analysis of the two genes,designated as cgi-smad1/5/8 and cgi-smad4,revealed conserved functional characteristics.The two genes were widely expressed in embryos and larvae,suggesting multiple roles in the early development of C.gigas.The mRNA of the two genes aggregated in the D quadrant and cgi-smad4 was highly expressed on the dorsal side of the gastrula,indicating that TGF-P signaling pathways may be involved in dorsoventral patterning in C.gigas.Furthermore,high expression levels of the two genes in the shell fields of embryos at different stages suggested important roles for TGF-P signaling pathways in particular phases of shell development,including the formation of the initial shell field and the biomineralization of larval shells.The results of this study provide fundamental support for elucidating how TGF-P signaling pathways participate in the early development of bivalve mollusks,and suggest that further work is warranted to this end.  相似文献   

12.
A feeding trial was conducted for 75 d to evaluate the nutritive value of a mixture of animal by-products(MAB) as a possible protein source in diets for juvenile mangrove red snapper,Lutjanus argentimaculatus(mean initial body weight,30 g). Fish were fed one of five isonitrogenous diets(40% crude protein) replacing 0,25%(MAB25),50%(MAB50),75%(MAB75) and 100%(MAB100) of fish meal protein with similar percentages of MAB. The MAB consisted of 25% cow liver meal,20% leather meal,20% meat and bone meal,15% blood meal,10% APC(poultry feather meal),8% poultry manure dried,1.5% choline and 0.5% chromic oxide. After 75 d of feeding,fish fed with diets MAB50,MAB75 and MAB100 exhibited significantly lower growth per-formance than that of fish fed with control and MAB25 diets. The optimum level of MAB was estimated to be 23%. Replacement of fish meal by MAB23% showed the following performance:maximum weight gain,510%;SGR,2.39% and FCE,2.83%. The MAB substitution up to 75% of fish meal protein in diets did not show differences in apparent protein digestibility(83.6% for MAB25,79.2% for MAB50,78.7% for MAB75) compared with control(83.4%),whereas in MAB100 group digestibility(65.3%) was sig-nificantly lower than in other groups. The apparent phosphorus absorption of test diet groups was significantly higher(37.1% for MAB25,28.5% for MAB50,55.6% for MAB75 and 54.5% for MAB100) than that of control(11.2%). The levels of protein and ash in the whole body,carcass and viscera increased as MAB substitution in diets increased,whereas lipids and moisture remained con-sistent among all treatment groups. These results showed that approximately 23% of fish meal protein could be replaced by a mixture of animal by-products for juvenile snapper growing from 30 g to 167 g in 75 d without compromising growth performance and feed efficiency.  相似文献   

13.
The effects of different dietary protein and lipid levels on juvenile greenfin horse-faced filefish(Thamnaconus septentrionalis) was experimentally evaluated in a 56-day study. Fish(initial weight 6.47 ± 0.56 g) were fed with nine different diet combinations containing three varying levels of protein(37%, 43%, 49%) and lipids(4%, 7%, 10%) in triplicates, where the protein-to-energy(P/E) ratios of the diets varied in the range of 19.17–26.99 mg kJ-1. Results showed that fish fed diets containing 49%protein and 10% lipid exhibited significantly lower feed intake than those fed other diets. The weight gain of fish fed diets with 43%protein was significantly higher than that of fish fed 37% or 49% protein diets. While no significant change in weight gain was observed when lipid content was increased from 4% to 7%, weight gain decreased significantly when lipid increased from 7% to 10%.The protein efficiency ratio and feed efficiency were highest when the fish were fed the diet with 49% protein and 7% lipids. Muscle crude lipid content was positively correlated with dietary protein level regardless of lipid level. The diet of 43% protein or 7% lipids significantly reduced the activity of alanine aminotransferase and aspartate aminotransferase in liver. The contents of aspartate aminotransferase, superoxide dismutase and malondialdehyde were significantly increased from 4% to 10% lipids in plasma. The plasma total cholesterol and triglyceride contents were positively correlated with dietary lipid level at the same protein level. Our comprehensive analysis concluded that a diet with 43% protein and 7% lipid, with a P/E ratio of 23.07 mg kJ-1, significantly promote the growth performance, feed utilization and health of juvenile greenfin horse-faced filefish.  相似文献   

14.
Understanding aquatic pathogen in sediments or aquacultural water is crucial to protect public health from soilborne and waterborne diseases.Quorum sensing (QS) was increasingly reported in biological wastewater treatment processes because of their inherent roles in biofilm development,bacterial aggregation and so on.The widely QS signals was Antoinducer-2 (AI-2),primarily involved to allow the possibility of interspecies communication.However,the cellular components that mediate the response of Streptococcus agalactiae to AI-2 have not been fully characterized.Analysis of the complete genome sequence of S.agalactiae indicated that its Rbs B protein has similarity to Escherichia coli Lsr B and Aggregatibacter actinomycetemcomitans Rbs B proteins that bind AI-2.We hypothesized that Rbs B protein mediates quorum sensing signal uptake via interaction with AI-2.To evaluate the regulatory effect of Rbs B on QS system,the recombinant plasmid p GEX-6p-1-Rbs B was constructed and Rbs B protein was purified with GST-tag.To further elucidate the role of Rbs B protein binding to DPD (AI-2 precursor dihydroxypentanedione),the systematically throughput circular dichroism (CD) spectroscopy,isothermal titration calorimetry_(200) (ITC_(200)) and molecular docking methods were employed.The high expression of soluble Rbs B protein with molecular weight of 33 k Da was obtained.The thermodynamics results (ΔH0,ΔS0,ΔG0) with ITC determination indicated that the binding process between DPD and Rbs B was exothermic and spontaneous,with hydrogen bonds and van der Waals forces as the main binding forces.Obviously,DPD can be more easily combined with Rbs B in a dose-dependent manner,suggesting that Rbs B was changed in the microenvironment of DPD when the DPD concentration was between 0.8–1.0 mmol L~(-1) and reaching the maximum binding amount.According to molecular docking,3hydrophobic residues involved in DPD and Rbs B protein stable binding were be found,and also hydrogen bonding plays a key role in the formation of the new complex.Rbs B efficiently inhibited V.harveyi bioluminescence induced by both S.agalactiae AI-2 and V.harveyi AI-2 in a dose-dependent manner.However,our results suggest that Rbs B may play a role in the response of S.agalactiace to AI-2.  相似文献   

15.
We investigated the effects of lipopolysaccharide(LPS) and dopamine(DA) on the activation of the prophenoloxidase(proPO) system of Litopenaeus vannamei.LPS and DA were shown with a negative dose-dependent effect on hyalne cells(HC),semi-granular cells(SGC),large granular cells(LGC),and total haemocyte count(THC).When haemocytes were treated with LPS or DA,serine proteinase activity and intracellular phenoloxidase(PO) activity were significantly reduced,but extracellular PO activity increased significantly.These findings indicated that the reduction in haemocyte counts was mainly because of the degranulation and activation of the proPO system from semi-granule and large granule cells.The PKC inhibitor,chelerythrine,and the TPK inhibitor,genistein,had an inhibitory effect on extracellular PO activity,while serine proteinase and intracellular PO activity increased.This suggests that the LPS and DA induce the activation of proPO in haemocytes via PKC and TPK-related signaling pathways,but serine proteinase may be activated only by PKC,as the genistein effects were not statistically significant.Electrophoresis analysis revealed that POs induced by LPS or DA have the same molecular mass and high diphenolase activity.Two PO bands at 526 kDa and 272 kDa were observed in PAGE,while in the haemocyte lysate supernatant(HLS),only a 272-kDa band was observed.This band was resolved after SDS-PAGE under non-reducing and reducing conditions into two groups of POs,166 kDa and 126 kDa,and 78.1 kDa and 73.6 kDa,respectively,suggesting that PO in L.vannamei is an oligomer,which may have different compositions intra-and extracellularly.  相似文献   

16.
An 8-week feeding trial was conducted to determine the optimal dietary protein-to-lipid ratio for juvenile Nibea albiflora with an initial weight of(11.76 ± 0.20) g.Nine experimental diets containing different concentrations of protein(40%,47%,or 54%) and lipids(5%,9%,or 13%) in a 3 × 3 factorial experimental design were tested in triplicate groups of fish,while the protein-to-energy(P/E) ratios of the diets varied in the range of 19.74–28.32 mg k J~(-1).Results showed that fish fed diets containing 9% or 13% lipids with 54% protein exhibited significantly higher weight gains and specific growth rates than those fed other diets.The feed conversion rate of fish fed the diet with 40% protein and 5% lipids was significantly poorer than that of fish fed other diets.The protein efficiency rate of fish fed diets with 5% lipids was significantly lower than that of fish fed 9% or 13% lipid diets.Carcass lipid and energy contents were positively correlated with dietary lipid level regardless of protein level.Fish fed a 54% protein diet showed the highest trypsin activity.The intestinal lipase activity of fish fed the diet containing 13% lipids was significantly higher than that of fish fed 5% or 9% lipid diets.These results demonstrate the high protein dietary requirements of N.albiflora.A diet containing 54% protein and 9%–13% lipids with a P/E ratio of 26.2–27.81 mg protein k J~(-1) can be considered optimal for juvenile N.albiflora.  相似文献   

17.
A 9-week feeding trial was conducted to investigate the effects of dietary carbohydrate to lipid ratio(CHO:LIP) on the growth performance and feed utilization of juvenile turbot Scophthalmus maximus(initial body weight 8.75 g ± 0.04 g). Four isonitrogenous and isoenergetic low protein level(39%) diets were formulated with increasing ratios of dietary carbohydrate to lipid(2:18, 6:18, 18:12 and 28:6). A high protein level(50%) diet with the 2:12 ratio of carbohydrate to lipid was used as the control. Results showed that the survival rate, contents of moisture, crude protein and ash in muscle were not significantly affected by dietary treatments. With the dietary CHO:LIP ratio increased from 2:18 to 18:12, weight gain rate significantly increased(P 0.05). Higher dietary CHO:LIP ratio(28:6) resulted in the significantly decreased weight gain rate(P 0.05). Meanwhile, this treatment also resulted in the highest daily feed intake and liver glycogen content, as well as the lowest feed efficiency(P 0.05). Muscle glycogen content in fish fed the diet with 2:12 or 2:18 CHO:LIP ratio was significantly lower than those fed with the other three diets(P 0.05). The present results confirmed that the juvenile turbot can utilize carbohydrate. Furthermore, the appropriate ratio of dietary carbohydrate to lipid was important to the growth and feed utilization of turbot. The proper CHO:LIP ratio based on the growth performance in the present study was determined to be 18:12 when the dietary protein level was 39%.  相似文献   

18.
Protein kinase C(PKC) has a crucial role in signal transduction for a variety of biologically active substances which activate cellular functions and proliferation. We previously isolated the full-length PKC gene from Dunaliella salina(Ds PKC) using rapid amplification of cDNA ends(RACE) and RT-PCR methods. And we submitted the mRNA sequence of Ds PKC gene to NCBI(Genbank No. JN625213). In the present paper, the Ds PKC gene open reading frame obtained by PCR was cloned into pGS-21 a vector and transformed into Escherichia coli to generate the fusion protein. Bioinformatics analysis revealed that DsPKC gene was a member of serine/threonine kinase with two conserved domains and highly conserved motifs. The Ds PKC was highly expressed upon induction with isopropyl-β-d-thiogalactoside(IPTG) at a final concentration of 0.2 mmol L~(-1) at 37℃. Under salt stress, the fusion protein Green Fluorescent Protein(GFP)-DsPKC was transferred from the cytoplasm to the cell membrane. The expression pattern of Ds PKC gene was analyzed using real-time quantitative PCR, and indicated that Ds PKC gene was up-regulated by 3.0 mol L~(-1) NaCl at 12 h, which was significantly higher than in control values(P 0.05). These results suggest that the Ds PKC gene plays an important role in response to salt stress in D. salina.  相似文献   

19.
Poor palatability is a limiting factor for replacing fishmeal with other protein sources in aquaculture. The water-soluble molecules with low molecular weights are the major determinants of the palatability of diets. The present study was conducted to investigate the palatability of water-soluble extracts from single protein source(single extract pellets) and the mixture of these extracts with different proportions(blended extract pellets) in juvenile turbot(Scophthalmus maximus). Then according to the palatability of blended extract pellets, an optimal mixture proportion was selected, and a new protein source made from raw protein materials with the selected proportion was formulated to replace fishmeal. Summarily, the palatability of single extract pellets for turbot was descendent from fishmeal to pet-food grade poultry by-product meal, wheat gluten meal, soybean meal, peanut meal, meat and bone meal, and corn gluten meal. Subsequently, according to the palatability of single extract pellets, 52 kinds of blended extract pellets were designed to test their palatability. The results showed that the pellets presented remarkably different palatability, and the optimal one was diet 52(wheat gluten meal: pet-food grade poultry by-product meal: meat and bone meal: corn gluten meal = 1:6:1:2). The highest ingestion ratio(the number of pellets ingested/the number of pellets fed) was 0.73 ± 0.03, which was observed in Diet 52. Then five isonitrogenous(52% crude protein) and isocaloric(20 k J g~(-1) gross energy) diets were formulated by replacing 0(control), 35%, 50%, 65% and 80% of fishmeal with No.52 blending proportion. After a 10-weeks feeding trial, a consistent feed intake was found among all replacement treatments. Replacement level of fishmeal up to 35% did not significantly influence final body weight, specific growth rate, feed efficiency ratio, and protein efficiency ratio of turbot. Therefore, the water-soluble extracts of protein sources play an important role in improving the palatability of non-fishmeal protein sources in aquafeed.  相似文献   

20.
Antarctic krill is a potentially nutritious food source for humans, but fluorine(F) toxicity is a matter of concern. To evaluate the toxicity of F in Antarctic krill, 30 Wistar rats were divided into three groups with different dietary regimens: a control group, a krill treatment group(150 mg·kg-1 F), and a sodium fluoride(NaF) treatment group(150 mg·kg-1 F). After three months, F concentrations in feces, plasma, and bone were determined, and the degree of dental and skeletal fluorosis was assessed. The F concentrations in plasma and bone from the krill treatment group were 0.167 0±0.020 4 mg.L-1 and 2 709.8±301.9 mg·kg-1, respectively, compared with 0.043 8±0.005 5 mg·L-1 and 442.4±60.7 mg·kg-1, respectively, in samples from the control group. Concentrations of F in plasma and bone in the krill treatment group were higher than in the control group, but lower than in the NaF treatment group. The degree of dental fluorosis in the krill treatment group was moderate, compared with severe in the NaF treatment group and normal in the control group. The degree of skeletal fluorosis did not change significantly in any group. These results showed that the toxicity of F in Antarctic krill was lower than for an equivalent concentration of F in NaF, but it was toxic for rats consuming krill in large quantities. To conclude, we discuss possible reasons for the reduced toxicity of F in Antarctic krill. The present study provides a direct toxicological reference for the consideration of Antarctic krill for human consumption.  相似文献   

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