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1.
Immunostimulants may improve disease resistance of aquaculture animals by promoting the nonspecific immunity response of the organisms. Five types of saccharides, including chitosan, yeast polysaccharide, burdock oligosaccharide, seaweed polysaccharide and lentinus edodes polysaccharide, were screened for potential use as immunostimulants by using spectrophotometry. The saccharides were injected into Apostichopus japonicus, a sea cucumber, and the lysozyme and superoxide dismutase (SOD) activities of the coelomic fluid and epidermal slime were monitored in six consecutive days. The results show that the lysozyme activity of the animal’s coelomic fluid was significantly stimulated on day 2, day 4 and day 6 after the injection of the saccharides (P<0.05). The effects of chitosan and yeast polysaccharide were the most notable. The lysozyme activity of the epidermal slime was significantly increased by chitosana, yeast polysaccharide, seaweed polysaccharide, and burdock oligosaccharide on day 1 and day 2 after the injection (P<0.05). The SOD activity of the coelomic fluid was significantly promoted by the saccharides on day 2 and day 4 post-injection (P<0.05), while the SOD activity of the epidermal slime increased on day 2. These findings indicate that chitosan and yeast polysaccharide are the most effective immunostimulants and potential healthy anti-disease feedstuff for A. japonicus. Supported by the National Key Technology Research and Development Program (863 Program, No. 2006BAD09A06) and the Special Fund of Chinese Central Government for Basic Scientific Research Operations in Commonweal Research Institutes (No. 02-2007B03)  相似文献   

2.
Viable cell count was used to determine whether Metschnikowia sp.C14 can colonize the intestine of juvenile sea cucumber Apostichopus japonicus.Sea cucumber individuals were divided into two groups,which were fed the control diet for 38 days or the C14-supplemented diet at 105 cells g−1 diet for 28 days,then the control diet from day 29 to day 38.The number of C14 cells in the intestine of sea cucumber fed the C14-supplemented diet significantly increased from day 7 to day 28,and decreased from day 29 to day 38.Sea cucumber fed with the diet containing C14 showed a significant increase in trypsin activity and lipase activity from day 21 to day 33 compared with the control.Feeding C14 significantly improved the phagocytic activity and respiratory burst in coelomocytes from day 21 to day 35 and from day 14 to day 38,respectively.In addition,there was an obvious enhancement in lysozyme activity(from day 21 to day 38 or day 33),phenoloxidase activity(from day 21 to day 28)and total nitric oxide synthase activity(from day 14 to day 38)in coelomic fluid supernatant and/or coelomocyte cell lysate supernatant compared with the control.There were significant positive correlations between the number of C14 cells colonizing the intestine and trypsin activity of the intestine,lysozyme activity of the coelomic fluid supernatant and coelomocyte lysate supernatant from sea cucumber.These data suggested that the number of C14 cells should be maintained at 105 cfu(colony-forming units)g−1 intestine material for the maximum benefit.  相似文献   

3.
Skin ulceration syndrome(SUS) is the main diseases affected the development of sea cucumber(Apostichopus japonicus) culture industries.To better observe the changes in the sea cucumber A.japonicus with SUS and understand the pathogenesis of the disease,activities of superoxide dismutase(SOD),catalase(CAT),and level of malondialdehyde(MDA) in coelomic fluid were detected using the Assay Kit and metabolite s in the body wall were assessed using ultra-performance liquid chromatography and quadrupoletime of flight mass spectrometry.The re sults indicated that level of MDA was increased during SUS compared with healthy individuals(P0.01),but activities of SOD and CAT were reduced(P0.05).In metabolomics analysis,metabolites,such as adenosine,choline,betaine aldehyde,palmitic acid,and taurine,were found to be upregulated and 2-oxoadipic acid,anthranilic acid(vitamin L1),thioetheramide-PC,cholesterol-3-sulfate,and pentadecanoic acid were downregulated(VIP1 and P0.1).Pathway enrichment analysis indicated most enrichment of KEGG pathways were mainly related to energy metabolism,immunity,and osmoregulation such as ABC transporters,glycine,serine and threonine metabolism,tryptophan metabolism and neuroactive ligand-receptor interaction.Our study reflected the difference in enzyme activity and metabolites between A.japonicus with SUS and those without,which will provide reference data for investigating SUS.  相似文献   

4.
A marine bacterium, Pseudoalteromonas sp. BC228 was supplemented to feed in a feeding experiment aiming to determine its ability of enhancing the digestive enzyme activity and immune response of juvenile Apostichopus japonicus. Sea cucumber individuals were fed with the diets containing 0(control), 105, 107 and 109 CFU g-1 diet of BC228 for 45 days. Results showed that intestinal trypsin and lipase activities were significantly enhanced by 107 and 109 CFU g-1 diet of BC228 in comparison with control(P 0.01). The phagocytic activity in the coelomocytes of sea cucumber fed the diet supplemented with 107 CFU g-1 diet of BC228 was significantly higher than that of those fed control diet(P 0.05). In addition, 105 and 107 CFU g-1 diet of BC228 significantly enhanced lysozyme and phenoloxidase activities in the coelomic fluid of sea cucumber, respectively, in comparison with other diets(P 0.01). Sea cucumbers, 10 each diet, were challenged with Vibrio splendidus NB13 after 45 days of feeding. It was found that the cumulative incidence and mortality of sea cucumber fed with BC228 containing diets were lower than those of animals fed control diet. Our findings evidenced that BC228 supplemented in diets improved the digestive enzyme activity of juvenile sea cucumber, stimulated its immune response and enhanced its resistance to the infection of V. splendidus.  相似文献   

5.
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6.
Grading procedure in routine sea cucumber hatchery production is thought to affect juvenile sea cucumber immunological response. The present study investigated the impact of a 3-min mechanical perturbation mimicking the grading procedure on neuroendocrine and immune parameters of the sea cucumber Apostichopus japonicus. During the application of stress, concentrations of noradrenaline and dopamine in coelomic fluid increased significantly, indicating that the mechanical perturbation resulted in a transient state of stress in sea cucumbers. Coelomocytes concentration in coelomic fluid increased transiently after the beginning of stressing, and reached the maximum in 1 h. Whereas, coelomocytes phagocytosis at 3 min, superoxide anion production from 3 min to 0.5 h, acid phosphatase activity at 0.5 h, and phenoloxidase activity from 3 min to 0.5 h were all significantly down-regulated. All of the immune parameters recovered to baseline levels after the experiment was conducted for 8 h, and an immunostimulation occurred after the stress considering the phagocytosis and acid phosphatase activity. The results suggested that, as in other marine invertebrates, neuroendocrine/immune connections exist in sea cucumber A. japonicus. Mechanical stress can elicit a profound influence on sea cucumber neuroendocrine system. Neuroendocrine messengers act in turn to modulate the immunity functions. Therefore, these effects should be considered for developing better husbandry procedures.  相似文献   

7.
Sea cucumbers, Apostichopus japonicus Selenka, were fed diets containing non-immunostimulant (basal diet), 0.2% β-glucan and 0.02% glycyrrhizin in a recirculatory water system for 45 days, and subsequently challenged with Vibrio splendidus by injection at 1.0×108 cfu / sea cucumber for 15 days. Phagocytic capacity (PC), intracellular superoxide anion production (ISAP), lysozyme (LSZ) activity and superoxide dismutase (SOD) activity in the coelomic fluid were analyzed on the 0th, 5th, 10th and 15th days after injection. Results showed that after the 45-day feeding period, PC, ISAP, LSZ activity and SOD activity in sea cucumbers fed with dietary β-glucan or glycyrrhizin were significantly higher than in those fed with the basal diet. On the 5th day after infection, all the immune parameters examined in the sea cucumbers injected with V. splendidus decreased in value significantly. On the 15th day, PC, ISAP and LSZ activity returned to levels similar to those on the 0th day. For the sea cucumbers injected with saline, there were no significant differences in all the immune parameters examined and in the cumulative morbidity during the 15-day challenging trial. After injecting with V. splendidus, the cumulative morbidity of sea cucumbers fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin when challenged with V. splendidus challenged sea cucumber fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin. There was no significant difference in cumulative morbidity between the dietary β-glucan and glycyrrhizin treatments over time.  相似文献   

8.
Information regarding antioxidant enzymes in amphioxus remains lacking, and this study was carried out to examine the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in different genders and tissues of amphioxus Branchiostoma belcheri tsingtauense. Results show that (1) CuZn-SOD, CAT and GPX activities in the whole amphioxus B. belcheri tsing- tauense were basically at the same levels in male and female amphioxus, whereas both T-SOD and Mn-SOD activities in male amphioxus were significantly higher than that in the female (P<0.05); (2) The testis had significantly higher T-SOD and CuZn-SOD activities than the ovary (P<0.05); (3) CuZn-SOD activity was undetectable in the guts of male and female amphioxus; (4) For both male and female am- phioxus, the activities of CAT and GPX in the gonads including testis and ovary were the lowest (P<0.05) among the tissues examined; (5) The gut and gill had the same level GPX activities while the gut had a higher CAT activity; (6) There was no clear difference in CAT and GPX activities in the corresponding tissues between male and female amphioxus. The study on SOD, CAT and GPX activities in different genders and tissues of the protochordate provides data for future comparison of amphioxus antioxidant enzymes with those of invertebrates and vertebrates.  相似文献   

9.
The effect of a potential probiotic on the growth performance and immune response of sea cucumber(Apostichopus japonicus) was investigated. Bacillus baekryungensis YD13 isolated from sea cucumber culturing ponds was added to sea cucumber basal feed as a probiotic in different doses(0, the control; 1×104(YD134), 1×106(YD136) and 1×108(YD138) CFU g-1 of diet), and administered orally to A. japonicus(initial mean wet weight 5.44 g ± 0.17 g). The sea cucumbers were fed in 20 aquaria, 5 each treatment, for 60 d. At the end of growth trial, 20 sea cucumbers from each treatment were challenged with Vibrio splendidus. A. japonicus in YD134 and YD136 exhibited significantly better growth performance than control(P 0.05). Five non-specific immune parameters including lysozyme, acid phosphatase, alkaline phosphatase, superoxide dismutase and catalase in coelomic fluid were measured to evaluate the immune response of A. japonicus to the probiotics. Results showed that all parameters were significantly improved when YD11 was supplemented in the dose of 1×106 CFU g-1(P 0.05). The cumulative incidence and mortality after the Vibrio splendidus challenge decreased significantly in sea cucumbers of YD136. Accordingly, 1×106 CFU g-1 of YD13 in diet was recommended for the growth promotion and immune enhancement of A. japonicus.  相似文献   

10.
Fucoidan is such a polysaccharide that its hydroxies are easy to combine with lanthanons ion (CeIV) to form complex. This work obtained the complexes of three fucoidan oligosaccharides with different molecular weights F1(>5000), F2 (1000-5000) and F3(<1000) by hydrolyzing Oligosaccharide collagen with sulfuric acid. It is found that the fucoidan oligosaccharide F3 can form complex with more CeIV than F1 and F2. Hydrolyzing collagen with the complex was carried out to produce amino acid and peptides. All the three fucoidan oligosaccharide complexes with CeIV( F1, F2, F3) can catalyze by the artificial hydrolytic enzyme, and the activity of the complex of F3 is the highest.  相似文献   

11.
The toxic effects of benzo[α]pyrene (B[α]P) at different concentrations (0.1, 0.5, 1, 2.5 and 7.5 ugL-1) on scallop (Chla-mys farreri) immune system were studied. The results showed that B[a]P had significant toxic effects on the haemocyte counts, neu-tral red uptake, phagocytosis, bacteriolytic and antibacterial activity (P<0.05), while the seawater control and acetone control had no significant differences. The haemocyte counts, neutral red uptake, phagocytosis and bacteriolvtic activity in all B[α]P treatment groups as well as antibacterial activity, in groups of 0.5, 1, 2.5 and 7.5 ugL-1 B[α]P decreased significantly (P<0.05). Some of these indices tended to be stable on the sixth day and others on the ninth day, and the indices showed clear time- and concentra-tion-response to B[α]P. Bacteriolytic activity in 0.1ugL-1 B[α]P treatment group and antibacterial activity in 0.1 ugL-1 and 0.5 ugL-1 B[α]P treatment groups increased at the beginning of exposure and reached their peaks on day 1 and day 6, respectively. Following that, both activities decreased gradually and became stable after day 9. When all the indices reached stability, they were significantly lower than those in control group (P<0.05), except for antibacterial activity in 0.1 ugL-1 B[α]P treatment group (P<0.05). Thus, B[α]P has evident toxic effects on scallop immune system, which supports the view that a relationship exists between pollution and immu-nomodulation in aquatic organisms.  相似文献   

12.
Fucoidan is such a polysaccharide that its hydroxies are easy to combine with lanthanons ion (CeⅣ) to form complex. This work obtained the complexes of three fucoidan oligosaccharides with different molecular weights F1( ＞5 000), F2(1 000-5 000) and F3( ＜ 1 000) by hydrolyzing Oligosaccharide collagen with sulfuric acid. It is found that the fucoidan oligosaccharide F3 can form complex with more CeⅣ than F1 and F2. Hydrolyzing collagen with the complex was carried out to produce amino acid and peptides. All the three fucoidan oligosaccharide complexes with CeⅣ( F1, F2, F3) can catalyze by the artificial hydrolytic enzyme, and the activity of the complex of F3 is the highest.  相似文献   

13.
Analgesis and wound healing effect of chitosan and carboxymethyl chitosan on scalded rats were investigated. A II degree scald model was established in rats, which was subsequently treated with chitosan and carboxymethyl chitosan solution, respectively. The concentration of bradykinin and 5-hydroxytryptophan was detected by assaying enzyme-linked immunosorbent. Healing condition was observed and pathological sections were made to determine the healing effect of chitosan and carboxymethyl chitosan. Results showed that the concentration of bradykinin and 5-hydroxytryptophan peaked at the third hour post-wound in all groups, while the concentration of hydroxyproline peaked at the seventh day post-wound in both chitosan and carboxymethyl chitosan group. The concentration of bradykinin and 5-hydroxytryptophan of carboxymethyl chitosan group was significantly lower than that of control (P < 0.05), while that of chitosan group was similar to that of control (P > 0.05). These findings indicated that carboxymethyl chitosan reduced the concentration of algogenic substances, resulting in analgesia. During the whole recovery process, the hydroxyproline concentration in chitosan and carboxymethyl chitosan group on day 3 and 7 was significantly higher than that of control (P < 0.01); however the significance of such a highness decreased on day 14 (P < 0.05). These findings indicated that chitosan and carboxymethyl chitosan accelerated tissue repair. Meanwhile, chitosan performed better in healing than carboxymethyl chitosan in both decrustation and healing time. In conclusion, carboxymethyl chitosan showed significant analgesis and wound-healing promotion effect, but chitosan only showed wound-healing promotion effect.  相似文献   

14.
Optimal stocking densities were investigated for the sea cucumber Apostichopus japonicus Selenka under feed-supplement and non-feed-supplement regimes in net enclosures for 333 d. Substantial weight loss occurred during the aestivation phase (AE).Decreased growth rates were also observed during the winter phase (WT). In contrast, sea cucumbers showed rapid growth during the spring (SP) and autumn (AU) phases. Feeding regimes considerably influenced the growth performance, i.e., sea cucumbers grew faster under feed-supplement regime than under non-feed-supplement regime (P< 0.05). The average survival rates of sea cucumbers under feed-supplement regime were higher than those under non-feed-supplement regime for both the autumn phase and spring phase,but the differences were only significant for the latter phase (P<0.05). The fitted B-N curves showed that the optimal stocking densities, in terms of net production, were 22.3 ind. m-2 for feed-supplement regime and 14.1 ind. m-2 for non-feed-supplement regime.  相似文献   

15.
The growth, metabolism and physiological response of the sea cucumber, Apostichopus japonicus, were investigated during periods of inactivity. The body weight, oxygen consumption rate (OCR), activities of acidic phosphatase (ACP), alkaline phosphatase (AKP), catalase (CAT) and superoxide dismutase (SOD), and content of heat shock protein 70 (Hsp70) in the body wall and coelomic fluid of A. japonicus were measured during starvation, experimental aestivation and aestivation. The results showed that the body weight of sea cucumber in the three treatments decreased significantly during the experimental period (P < 0.05). The OCR of sea cucumber reduced in starvation and experimental aestivation treatments, but increased gradually in natural aestivation treatment. The activities of ACP and AKP of sea cucumber decreased gradually in all treatments, whereas those of SOD and CAT as well as Hsp70 content decreased in the starvation and experimental aestivation treatments and increased in natural aestivation treatment. The sea cucumber entered a state of aestivation at 24℃. To some extent, the animals in experimental aestivation were different from those in natural aestivation in metabolism and physiological response. These findings suggested that the aestivation mechanism of A. japonicus is complex and may not be attributed to the elevated temperature only.  相似文献   

16.
To investigate the stimulating effects of immunostimulants on the autogenous immunocompetence of crabs and the possible mechanisms involved, the immunostimulating effects of β-1,3-glucan, lipopolysaccharide (LPS), inactivated Vibrio harveyi and Vibrio anguillarum on phenoloxidase (PO) and hemocytes of Charybdis japonica were investigated in this study. It was found that the yields and the enzymatic activities of purified PO in C. japonica increased significantly after the crabs were treated with immunostimulants, while the unit enzymatic activities remained almost the same. After treatment with β-1,3-glucan and LPS, the amount of rough endoplasmic reticulum (RER) and the number of mitochondria in both semigranular cells and granular cells increased greatly, and the number of cytoplasmic granules decreased but with enlarged volume. However, the corresponding characteristics of hyaline cells remained almost the same. On the other hand, the number of granules in semigranular cells decreased greatly, and the number of mitochondria of hyaline cells increased greatly, after treatment with inactivated vibrios. It may be concluded that the effect of polysaccharide immunostimulants on the innate immune system of C. japonica is different from that of inactivated vibrio immunostimulants. The immunity-enhancing mechanism of polysaccharides in crab autogenous immunocompetence is probably accomplished by the increased yields of PO and total PO activities, while that of inactivated vibrios is probably accomplished by the partially increased yields of PO and total PO activities as well as the significantly improved phagocytotic abilities of semigranular cells and hyaline cells.  相似文献   

17.
采用人工合成的10种未甲基化CpG-ODNs序列分别注射健康的吉富罗非鱼Oreochromis niloticus,48 h后注射海豚链球菌(Streptococcus iniae)进行人工感染,分别于感染24 h和48 h后检测鱼体血清溶菌酶(Lysozyme,LSZ)、超氧化物歧化酶(Superoxide dismutase,SOD)和碱性磷酸酶(Alkaline phosphatase,AKP)的活性,并记录死亡率。结果显示:注射ODN-205的罗非鱼LSZ、AKP和SOD活性变化均不显著(P>0.05),推测注射ODN-205后罗非鱼能较好的维持机体的平衡,增强机体的抗菌能力;注射ODN-1681、1670、2133和2006的罗非鱼LSZ活性逐渐降低,而注射ODN-1826、2143、1668、2102、1669的罗非鱼LSZ活性呈现先升高后降低的趋势。AKP活性的变化趋势同LSZ活性变化大体相似,而SOD活性前后变化不显著(p值>0.05)。注射CpG-ODNs的实验组抑制海豚链球菌感染的作用差异显著(p值<0.05),其中以注射ODN-205组的死亡率最低(18%),其次为注射ODN-1670、1681、2006和2133组,死亡率均低于30%,而ODN-1669组和生理盐水组的死亡率均超过90%。实验表明,注射一定量的ODN-205、1670、1681、2006和2133能明显提高吉富罗非鱼抗海豚链球菌感染能力。  相似文献   

18.
In nature, Apostichopus japonicus exhibits a behavioral characteristic of emerging at night and sheltering during the day. Hence, it was commonly believed that longer and darker light conditions are better for a population of A. japonicus. In aquaculture situations therefore, animals have commonly been cultured without lighting, especially during juvenile-hatching culture. However, how the length of darkness each day affects A. japonicus and what the mechanism is of behavioral response of A. japonicus to different photoperiods remain unclear, and are specifically addressed in this study. We applied nine photoperiod treatments (light:dark (LD) cycles at 0:24, 3:21, 6:18, 9:15, 12:12, 15:9, 18:6, 21:3, and 24:0 under 500 lx) to 60 individuals (29.73 ± 0.23 g) per treatment (with 3 replicates). The environmental conditions were set as: water temperature = 16.9 ± 0.3°C, dissolved oxygen>6.0 mg/L, ammonia<0.3 mg/L, pH = 7.8-8.2, and salinity = 30-32. Under experimental conditions of continuous darkness as well as continuous light, cyclic nocturnal activity patterns of A. japonicus (viz. the animals emerged and fed at night and sheltered during the day) were observed. However, they spent more and less time, respectively, moving and feeding under continuous darkness and continuous light, than those under a natural light cycle. Under photoperiods with 6-12 h of light, the animals showed one sheltering behavior transition and two emerging behavior transitions. These behavior transitions appeared to be governed by some internal physiological factors and induced by daily light variation. The behavior of the animals was significantly affected by different photoperiods, and the distribution rate (DR) of the animals emerging at "daytime" increased with lengthening "light time" (except for the LD 3:21 cycle). We also found that there was no significant difference in mean DR per day under LD cycles of 6:18, 9:15, 15:9, and 12:12. Mean DRs per day were significantly lower for these photoperiods, than for those under LD cycles of 0:24 and 3:21, and significantly higher than for those under LD cycles of 18:6, 21:3 and 24:0. These results imply that 6-15 h light per day were the similar photoperiods to natural sunlight. From the viewpoint of activity and feeding behaviors, these might be the optimal photoperiods for rearing A. japonicus.  相似文献   

19.
Sea cucumber A postichopus japonicus stock enhancement by releasing hatchery-produced seeds is a management tool used to recover its population under natural environmental conditions. To assess the suitability of releasing sites, we examined the microbiota of the gut contents of A. japonicus from two populations(one in sandy-muddy seagrass beds and one in rocky intertidal reefs) and the microbiota in their surrounding sediments. The activities of digestive and immune-related enzymes in the A. japonicus were also examined. The results indicated that higher bacterial richness and Shannon diversity index were observed in all the seagrass-bed samples. There were significant differences in intestinal and sediment microorganisms between the two habitats, with a 2.87 times higher abundance of Firmicutes in the seagrass bed sediments than that in the reefs. Meanwhile, Bacteroidetes and Actinobacteria were significantly higher abundant in the gut content of A. japonicus from seagrass bed than those from the reefs. In addition, the seagrass-bed samples exhibited a relatively higher abundance of potential probiotics. Principal coordinates analysis and heatmap showed the bacterial communities were classifi ed into two groups corresponding to the two habitat types. Moreover, compared to A. japonicus obtained from rocky intertidal habitat, those obtained from the seagrass bed showed higher lysozyme, superoxide dismutase and protease activities. Our results suggest that bacterial communities present in seagrass beds might enhance the digestive function and immunity of A. japonicus. Therefore, compared with the rocky intertidal reef, seagrass bed seems to be more beneficial for the survival of A. japonicus.  相似文献   

20.
采用中心组合设计和响应曲面法研究温度(12~34℃)和盐度(0~26)对吉富品系尼罗罗非鱼(Oreochromis niloticus)幼鱼(体长4.20±0.10 cm,体质量2.42±0.15 g)肠道超氧化物歧化酶(Superoxide Dismutase,SOD)、过氧化氢酶(Catalane,CAT)的联合效应。结果表明:1)温度对吉富罗非鱼幼鱼肠道SOD和CAT活力的线性效应和累积效应均极显著(P<0.01);盐度对吉富罗非鱼幼鱼肠道SOD活力线性效应显著(P<0.05),累积效应极显著(P<0.01);盐度对吉富罗非鱼幼鱼肠道CAT活力线性效应不显著(P>0.05),累积效应极显著(P<0.01);温度和盐度对SOD活力的互作效应极显著(P<0.01),对CAT活力的互作效应不显著(P>0.05)。2)响应曲面法分析表明,随着温度和盐度的增大,两种抗氧化酶的活力均呈先减小后增大的趋势。3)建立两种抗氧化酶活力与温度、盐度间关系的模型方程(SOD:决定系数R2=0.992 9,预测决定系数R2Pred.=0.939 2,P<0.01;CAT:决定系数R2=0.948 5,预测决定系数R2Pred.=0.647 1,P<0.01),两模型方程可用于预测吉富罗非鱼幼鱼肠道SOD和CAT的活力。4)优化结果显示,温度为24.15℃,盐度为11.75时,SOD活力最小值为9.80 U/mg,CAT活力最小值为9.28 U/mg,满意度函数值高达0.961。  相似文献   

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