Modulation by Biogenic Amines for the Hemocyte Count and Prophenoloxidase Exocytosis via Receptors in Litopenaeus vannamei

Hemocyte counts and phenoloxidase(PO)activity were examined after hemolymph being incubated in dopamine(DA),noradrenaline(NE)and serotonin(5-HT).Results showed that all the three biogenic amines(BAs)had a significant impact on total hemocyte count(THC),differential hemocyte count(DHC),and intracelluar and extracelluar phenoloxidase(PO)activity.Among these Bas,DA had the strongest effect on the above parameters,whereas 5-HT had the least effect.Preincubation with D1 receptor antagonist SCH23390,D2 receptor a...     

The roles of serine protease, intracellular and extracellular phenoloxidase in activation of prophenoloxidase system, and characterization of phenoloxidase from shrimp haemocytes induced by lipopolysaccharide or dopamine

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.     

Effect of dopamine injection on the hemocyte count and prophenoloxidase system of the white shrimp Litopenaeus vannamei

Effects of dopamine injection on the hemocyte count, phenoloxidase activity, serine proteinase activity, proteinase in-hibitor activity and α2-macroglobulin-like activity in L. vannamei were studied. Results showed that dopamine injection resulted in a significant effect on the parameters measured (P < 0.05), while no significant difference was observed in the control group (0.85% NaCl). In the experimental groups,the hemocyte count reached the minimum in 3 h;granular and semi-granular cells became stable after 12 h and hyaline cells and the total hemocyte count became stable after 18 h. Phenoloxidase activity reached the minimum in 6 h, and then became stable after 9 h. Serine protease activity and proteinase inhibitor activity reached the minimum in 3 h, and α2-macro-globulin-like activity reached the maximum in 3 h,and all the three parameters became stable after 12 h. The results suggest that the activating mechanisms of the proPO system triggered by dopamine are different from those triggered by invasive agents or sponta-neously activated under a normal physical condition.     

Effects of Hypoxia on Dopamine Concentration and the Immune Response of White Shrimp （Litopenaeus vannamei）

Effects of hypoxia on the dopamine concentration and the immune response of White Shrimp Litopenaeus vannamei were studied. The results showed that hypoxia had significant effects on the concentration of dopamine (DA) in the haemolymph, haemocyte count, phenoloxidase activity, phagocytic activity of haemocytes and bacteriolytic and antibacterial activity in the haemolvmph (P<0.05). The concentration of the dopamine in haemolymph reached its maximum in the 3.0 and 1.5mgL-1 DO groups at 12h and 6h, and then returned to normal after 24h and 12h, respectively. All immune parameters decreased with the reduction of dissolved oxygen. Total haemocyte count (THC), the hyaline cells and semi-granular cells in the 3.0mgL-1 DO group became stable after 12h,while granular cells did so after 24h, The THC and different haemocyte count (DHC) in the 1.5mgL-1 DO group became stable alter 24h. Phenoloxidase activity and bacteriolytic activity in the 3.0 and 1.5mgL-1 DO groups reached their stable levels after 24 h and 12 h respectively, while phagocytic activity and antibacterial activity became stable after 24 and 12, and 36 and 24 It, respec-tively. It was also indicated that the changes of dopamine concentrations in haemolymph, haemocyte count and phenoloxidase activ-ity were obviously related to the exposure time under hypoxic conditions.     

Effects of hypoxia on dopamine concentration and the immune response of White Shrimp (Litopenaeus vannamei)

Effects of hypoxia on the dopamine concentration and the immune response of White Shrimp Litopenaeus vannamei were studied. The results showed that hypoxia had significant effects on the concentration of dopamine (DA) in the haemolymph, haemocyte count, phenoloxidase activity, phagocytic activity of haemocytes and bacteriolytic and antibacterial activity in the haemolymph (P<0.05). The concentration of the dopamine in haemolymph reached its maximum in the 3.0 and 1.5 mg L⁻¹ DO groups at 12 h and 6 h, and then returned to normal after 24 h and 12 h, respectively. All immune parameters decreased with the reduction of dissolved oxygen. Total haemocyte count (THC), the hyaline cells and semi-granular cells in the 3.0 mg L⁻¹ DO group became stable after 12 h, while granular cells did so after 24 h. The THC and different haemocyte count (DHC) in the 1.5 mg L⁻¹ DO group became stable after 24 h. Phenoloxidase activity and bacteriolytic activity in the 3.0 and 1.5 mg L⁻¹ DO groups reached their stable levels after 24 h and 12 h respectively, while phagocytic activity and antibacterial activity became stable after 24 and 12, and 36 and 24 h, respectively. It was also indicated that the changes of dopamine concentrations in haemolymph, haemocyte count and phenoloxidase activity were obviously related to the exposure time under hypoxic conditions.     

Responses of prophenoloxidase system and related defence parameters of <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> to low salinity

In this study, we investigated the effects of low salinity(26 and 21)on the prophenoloxidase(proPO)system and related defence parameters in the shrimp Litopenaeus vannamei. The results showed that low salinity induced a significant increase of dopamine(DA)concentration in haemolymph at 6h of the experiment; on the other hand, total haemocyte count(THC), differential haemocyte count(DHC)and PO activity decreased over time to the lowest level at 24 h and remained low thereafter. Serine Protease(SP)and Proteinase Inhibitor(PI)activity in the two lower salinity treatments decreased to the lowest level at 12 and 24h, respectively, and both recovered to the control level at 72 h. In contrast, α2-macroglobulin(α2M)activity in the two lower salinity treatments peaked at 24h and then decreased to the control level at 72h. Therefore, it may be concluded that stress-induced DA plays an important temporary role in neurotransmission and causes immune response in L. Vannamei in adapting to salinity changes.     

Study on biogenic amines in various dry salted fish consumed in China

This study was carried out to investigate the biogenic amines（BAs）, physicochemical property and microorganisms in dry salted fish, a traditional aquatic food consumed in China. Forty three samples of dry salted fish were gathered from retail and wholesale markets and manufacturers, which had been produced in various regions in China. Cadaverine（CAD） and putrescine（PUT） were quantitatively the most common biogenic amines. About 14% of the samples exceeded the histamine content standards established by the FDA and/or EU. The highest histamine content was found in Silver pomfret（Pampus argenteus）(347.79 mg kg^-1). Five of forty three samples exceeded the acceptable content of TYR(100 mg kg^-1), and 23.26% of dried-salted fish contained high contents of biogenic amines(above 600 mg kg^-1). In addition, species, regions, pickling processes and drying methods made the physicochemical property, microorganisms and biogenic amines in dry salted fish to be different to some extents. The total plate count（TPC） was much higher than that of total halophilic bacteria in all samples. The biogenic amines, physicochemical property and microbiological counts exhibited large variations among samples. Furthermore, no significant correlation between biogenic amines and physicochemical property and TPC was observed. This study indicated that dry salted fish may still present healthy risk for BAs, depending on the processing methods, storage conditions among others.     

A Comparative study on the nonspecific immunity of juvenile Litopenaeus vannamei ever inhabiting freshwater and seawater

A study on the nonspecific immunity of Litopenaeus vannamei ever inhabiting freshwater and seawater was carried out at different molt stages by comparing their total hemocyte count(THC) and respiratory burst(RB) and activity of phenol oxidase(PO), nitric oxide synthase(NOS) and lysozyme(LY). Two-way ANOVA showed that salinity and molt stage independently affected THC and RB and the activity of PO, NOS and LY of juvenile L. vannamei significantly(P 0.05). The THC and RB and the activity of NOS gradually increased from the post-molt stages(A and B) to the pre-molt stages(D0–D3), which were common in shrimps inhabiting freshwater and seawater. The activity of PO peaked at the inter-molt stage(C), and touched the lowest at the post-molt stage in freshwater and pre-molt stage in seawater. The activity of LY was stable over the molt cycle. The RB and the activity of PO, NOS and LY of juvenile L. vannamei were significantly lower in freshwater than in seawater; whereas THC was significantly higher in freshwater than in seawater(P 0.05). It was concluded that the post-molt stage(especially stage A) was critical to shrimp culture, which should be intensively attended when L. vannamei was cultured in freshwater.     

Effects of Several Immunostimulants on Phenoloxidase and Hemocytes of the Crab Charybdis japonica

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.     

Effects of several immunostimulants on phenoloxidase and hemocytes of the crab Charybdis japonica

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.     

Dietary guaiacol improves the growth of juvenile abalone, <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> Ino

We conducted a preliminarily study on the effects of dietary guaiacol on growth performance of abalone, Haliotis discus hannai Ino. Seven semi-purified diets were formulated with graded levels of guaiacol (0, 5, 10, 50, 100, 500, and 1 000 mg kg⁻¹). Abalone (initial weight: 0.29 ± 0.01 g; initial shell length: 8.55 ± 0.27 mm) were fed with these diets in a re-circulated water system for 152 days. Guaiacol significantly improved the specific growth rate (SGR) (P<0.05); excessive dietary guaiacol (1 000 mg kg⁻¹) led to significantly high mortality (P<0.05), and lipid content in the soft body increased significantly after dietary guaiacol (P<0.05). Activities of catalase and phenoloxidase (PO) in the viscera were significantly stimulated by dietary guaiacol (P<0.05). Broken-line analysis based on SGR indicated that the minimum dietary guaiacol for the optimal growth of juvenile abalone is 15.43 mg kg⁻¹.     

全球海洋初级生产力时空异常变化对ENSO事件的响应

海洋初级生产力在海洋环境要素的驱动下,在不同海域呈现出不同的时空变化特征,这种时空演变特征在不同的ENSO事件类型下差异更为显著。本文基于1998年1月至2017年12月全球海洋初级生产力的卫星遥感数据集,通过改进海洋时空双约束聚类挖掘方法,挖掘了近20年海洋初级生产力的时空聚簇模式,并从时空分布和空间移动2个方面对比分析了海洋初级生产力时空演变簇与ENSO（El Niño-Southern Oscillation）事件之间的关系。结果表明：① 在EP（Eastern-Pacific）型El Niño事件期间,海洋初级生产力异常低值时空簇主要分布在赤道太平洋东部或中东部海域,异常高值时空簇主要分布在西太平洋和南太平洋中部海域;在CP（Central-Pacific）型El Niño事件期间,异常低值时空簇分布在太平洋中部,而异常高值时空簇分布在南太平洋与西太平洋海域;② 在EP型La Niña事件期间,赤道太平洋中部及东部、赤道大西洋与印度洋海域出现异常高值时空簇,南太平洋中东部海域出现异常低值时空簇;在CP型La Niña事件期间,赤道太平洋中部出现异常高值时空簇;南太平洋中西部海域出现异常低值时空簇;③ 发生在赤道太平洋的海洋初级生产力时空演变簇,在EP型ENSO事件期间具有东移特征,而在CP型ENSO事件期间,时空演变簇在赤道太平洋中部海域产生并消亡;④ ENSO事件中海洋初级生产力时空演变簇面积与MEI具有较强相关性。     

桂西南喀斯特-北部湾海岸带生态环境脆弱性时空分异与驱动机制研究

为揭示生态环境脆弱性的时空分异和驱动因子,本研究在山江海视角下,以桂西南喀斯特-北部湾海岸带为典型研究区,运用空间主成分分析法,地理探测器模型,结合生态环境脆弱性综合指数,系统分析桂西南喀斯特-北部湾海岸带生态环境脆弱性的时空分异特征及驱动机制。结果表明：① 研究区2008、2013、2018年脆弱性指数分别为0.54、0.61、0.69,多年平均值为0.61,整体处于中度脆弱,在空间上,由城市中心向四周逐渐降低的趋势;在时间上,生态环境脆弱等级呈微恶化趋势; ② 在单因子作用中6个驱动因子对生态环境脆弱性的解释力强度为汛期降雨量（0.457）>植被覆盖度（0.384）>高温季节温度（0.311）>废水入海量（0.248）>NPP（0.184）>人口密度（0.036）。在多因子交互中,只有汛期降水量和NPP, NPP和高温季节温度、废水入海量和NPP呈非线性增强,其余的交互作用均为双线性增强,而且汛期降水量和植被覆盖度的单因子影响较强,交互作用后影响也是最强（0.679）,说明了汛期降水量和植被覆盖度为该区域的主要驱动因子。     

GNSS UTCO参数演变分析与对比评估北大核心CSCD

全球导航卫星系统(GNSS)发播的协调世界时偏差参数(UTCO)随着导航系统现代化升级而更新,尤其是GPS和BDS的UTCO参数具有明显变化。从模型上看,一阶溯源模型改为二阶溯源模型,兼容一阶模型;从分辨率上看,参数分辨率普遍变小,从而提高对UTCO微小变化的敏感性。通过研究GPS、BDS、Galileo、GLONASS四大导航系统GPS LNAV电文、GPS CNAV电文、BDS D1/D2电文、BDS B-CNAV电文、Galileo电文、GLONASS电文中的UTCO参数,基于国际计量局(BIPM)国际时间公报(T公报)UTCr-UTC(k)数据,将GNSS UTCO参数统一到快速协调世界时(UTCr),分析不同系统发播的UTCO参数性能。同时以国家授时中心(NTSC)监测的GNSS系统时差和BIPM快速T公报作为参考,评估GNSS系统发播的协调世界时偏差误差(UTCOE)。结果表明,在实验时段内,GPS CNAV电文的UTCO参数性能优于GPS LNAV电文,BDS B-CNAV电文的UTCO参数性能优于BDS D1/D2电文;GPS、Galileo的UTCO参数性能优于BDS、GLONASS,BDS和GLONASS在UTC溯源中具有一定的系统误差。     

Effect of nitrogen to phosphorus ratios on cell proliferation in marine micro algae

The ratio of nitrogen/phosphorus (N/P) is known to affect cell proliferation of some marine micro algae. We evaluated the effect of N/P ratios on the proliferation and succession of phytoplankton using five marine micro algae species. We used two sources of nitrogen, NH4Cl (N1) and urea (N2), and a single source of phosphorous, NaH2PO4(P). The optimal N/P ratio that differed among the five species was affected by the source of nitrogen, being as follows (N1/P, N2/P in order): Thalassiosira sp. (30/1, 20/1),...     

Enhanced resistance of Portunus trituberculatus to Vibrio alginolyticus by selective breeding

We established a line (screened) of Portunus trituberculatus by selectively breeding individuals that survived from challenge with Vibrio alginolyticus, and compared the response of screened and unscreened (control) P. trituberculatus challenged with V. alginolyticus. We measured superoxide dismutase, catalase, acid phosphatase, alkaline phosphatase, and peroxidase activity and the content of hemocyanin in the plasma and phenoloxidase activity in serum. The cumulative survival rate after 24-h challenge with V. alginolyticus was significantly higher in the screened crabs than in the unscreened crabs (P<0.05). T-SOD and PO activity were significantly lower in the screened stock than in the unscreened stock (P<0.05). POD, CAT, and ACP activity and hemocyanin content were significantly higher in the screened stock than in the unscreened stock. Our results suggest that the screened stock was more resistant to infection. Furthermore, the indices we measured may be used to evaluate the health state of P. trituberculatus.     

Effects of different enzymatic hydrolysis methods on the bioactivity of peptidoglycan in Litopenaeus vannamei

The effects of different hydrolysis methods on peptidoglycan(PG) were assessed in terms of their impact on the innate immunity and disease resistance of Pacific white shrimp,Litopenaeus vannamei.PG derived from Bifidobacterium thermophilum was prepared in the laboratory and processed with lysozyme and protease under varying conditions to produce several different PG preparations.A standard shrimp feed was mixed with 0.05% PG preparations to produce a number of experimental diets for shrimp.The composition,concentration,and molecular weight ranges of the soluble PG were analyzed.Serum phenoloxidase and acid phosphatase activity in the shrimp were determined on Days 6-31 of the experiment.The protective activity of the PG preparations was evaluated by exposing shrimp to white spot syndrome virus(WSSV).Data on the composition of the PG preparations indicated that preparations hydrolyzed with lysozyme for 72 h had more low-molecular-weight PG than those treated for 24 h,and hydrolysis by protease enhanced efficiency of hydrolysis compared to lysozyme.SDS-PAGE showed changes in the molecular weight of the soluble PG produced by the different hydrolysis methods.Measurements of serum phenoloxidase and acid phosphatase activity levels in the shrimp indicated that the PG preparations processed with enzymes were superior to the preparation which had not undergone hydrolysis in enhancing the activity of the two serum enzymes.In addition,the preparation containing more low-molecular-weight PG enhanced the resistance of the shrimp to WSSV,whereas no increased resistance was observed for preparations containing less low-molecular-weight PG.These findings suggest that the immunity-enhancing activity of PG is related to its molecular weight and that increasing the quantity of low-molecular-weight PG can fortify the effect of immunity enhancement.