Enzymatic properties of UFE, a novel marine fibrinolytic enzyme

A novel potent protease, Urechis unicinctus fibrinolytic enzyme (UFE), was firstly discovered. The enzymatic properties of UFE were further investigated. As a low molecular mass protein, UFE appeared to be very stable to heat and pH. When temperature was below 50 ℃, the remnant enzyme activity remained almost unchanged, but when temperature was raised to 60 ℃, the remnant enzyme activity began to decrease rapidly. UFE was quite stable in the range of pH value from 3 to 12, especially in slightly alkaline pH value. Mn2 , Cu2 and Fe2 ions were activators of UFE, while Fe3 and Ag ions were inhibitors of UFE. Fe2 ion along with Fe3 ion might regulate UFE activity in vivo. The optimum pH and temperature of UFE were about 8 and 50 ℃, respectively. Other characteristics of this enzyme were also studied. Systematic research results are significant when UFE is applied for medical and industrial purposes.     

DDT对细巧华哲水蚤（Sinocalanus tenellus）摄食、呼吸、存活和生殖的影响研究

DDT(dichloro-diphenyl-trichloroethane) as a type of organochlorine pesticides, is an important component of pesticides pollution whose impact on the marine ecosystem is urgently to be evaluated. To investigate the biological effects of DDT on the marine ecosystem, copepods being the main contributor of secondary productivity in the marine ecosystem, were selected as target animals. The influence of DDT on the feeding, respiration, survival, and reproduction of Sinocalanus tenellus(S. tenellus) was analyzed and the antioxidant enzymes activities in the individuals were measured under different exposure concentrations of DDT. The 48 h median lethal concentration(LC50) and 96 h LC50 of DDT to S. tenellus were 5.44 and 2.50 μg/dm3, respectively. The filtration rates, grazing rates, and respiration of S. tenellus decreased apparently with increased DDT concentrations. Under lower concentration(625 ng/dm3) of DDT, the activities of the antioxidant enzymes, including superoxide dismutase and catalase in the animals increased significantly compared with those in the animals without any exposure to DDT, which suggested that the antioxidant enzymes can protect the animals from oxidative damage. However, the activity of the antioxidant enzyme decreased when the animals were exposed to higher concentration(1 250–2 500 ng/dm3) of DDT. The survival rate of both females and males was reduced when they were exposed to DDT less than 250 ng/dm3, but females showed higher survival rate than males when they are under the same concentration. The hatching ratio and the egg diameters of S. tenellus decreased significantly when they were exposed to DDT with a concentration of 25 and 250 ng/dm3, however, the cumulative egg production did not show any significant variation when the animals were exposed to the above DDT concentration. These data in the preset study suggested that exposure to DDT can cause the variation of the species composition of copepods, and further affect the marine ecosystem.     

Response of Yesso scallop Patinopecten yessoensis to acute temperature challenge: physiological and biochemical parameters

Water temperature is generally considered to be a major factor af fecting the physiological and biochemical activities of marine bivalves. Here, the physiological and biochemical responses of Yesso scallop, Patinopecten yessoensis, to acute water temperature changes in summer were studied. Scallops were transferred directly to a lower temperature( T dec treatment)(from 23°C to 15°C) or to a higher temperature( Ti nc treatment)(from 15°C to 23°C) for 72 h, respectively. Results showed that the oxygen consumption and ammonia-N excretion rates of P. yessoensis decreased signi?cantly in the T dec treatment but increased dramatically at 6 h in the Ti nc treatment( P <0.05). In the T dec treatment, hepatopancreas antioxidant enzyme activities, superoxide dismutase(SOD) and catalase(CAT) activities, increased substantially within 72 h( P <0.05). However, a signi?cant decrease in CAT activity was found at 12 h in the Ti nc treatment( P <0.01).A signi?cant enhancement of acid phosphatase(ACP) activity and malondialdehyde(MDA) content was detected when scallops were acutely exposed to a temperature of 15°C. The levels of Cu/Zn-SOD gene expression in their gills up-regulated signi?cantly in response to acute temperature changes( P <0.01). These data suggest that acute temperature change af fects physiological and biochemical functions, and improve our knowledge of P. yessoensis under conditions of thermal stress.     

Effect of oil pollution on gluthione and relative enzyme in oyster(Saccostrea cuculiata)

The oysters ( Saccostrea cuculiata ) were collected from four stations around Xiamen Island (Lundu Port, Xinglin Bay, Tong" an Bay, Huangcuo). The relation between the level of petroleum hydrocarbon in whole tissue and the contents of glutathione (GSH), the activity of selenium-dependent glutathione peroxidase (Se-GPx) and glutathione S-transferase (GST) in digestive gland and gill were investigated. The results showed: (1) The contents of petroleum hydrocarbon in oyster collected from four stations (Lundu Port, Xinglin Bay, Tong'an Bay, Huangcuo) were 380.68, 112.34, 27.31, 20.37μg/g wet weight, respectively; (2) the activity of Se-GPx and GST in digestive gland was lower than that in gill, and the content of GSH seemed reversibly; (3) among the four stations, both Se-GPx and GST activity of digestive gland and gill in Saccostrea cuculiata sampled from the four stations showed a good correlation with whole tissue petroleum hydrocarbon, could be as biomarkers of sea oil pollution.     

Comparison of copper speciation in the Zhujiang River Estuary between summer and winter:implication for upwelling current

The contents of total copper, total dissolved copper, particulate copper, and free ion copper in the Zhujiang River Estuary were investigated in July 2002 and January 2003 respectively, and the spatial distribution trend of these species of copper were compared in two cruises. It was found that, in summer, the total copper content in the Zhujiang River Estuary increased with salinity, while the content office ion copper decreased with salinity. However, in winter, the contents of total copper and free ion copper both decreased with salinity. So, it could be concluded that, copper content and speciation in the Zhujiang River Estuary were regulated by other factors than runoffdiffusion such as upwelling current. In order to prove the spatial trend of rice ion copper content in the Zhujiang River Estuary, the cultivation of Dunaliella tertiolecta in waters obtained from sampling stations with different salinities was carried out, and the copper uptake by algae cells was measured then. It was found that, the cells＇ uptake of copper increased with rice ion copper, not the content of total copper.     

深海嗜热菌Geobacillus sp. EPT3锰超氧化物歧化酶的纯化与生物化学鉴定

Thermostable SOD is a promising enzyme in biotechnological applications. In the present study, thermophile Geobacillus sp. EPT3 was isolated from a deep-sea hydrothermal field in the East Pacific. A thermostable superoxide dismutase(SOD) from this strain was purified to homogeneity by steps of fractional ammonium sulfate precipitation, DEAE-Sepharose chromatography, and Phenyl-Sepharose chromatography. SOD was purified 13.4 fold to homogeneity with a specific activity of 3 354 U/mg and 11.1% recovery. SOD from Geobacillus sp. EPT3 was of the Mn-SOD type, judged by the insensitivity of the enzyme to both KCN and H2O2. SOD was determined to be a homodimer with monomeric molecular mass of 26.0 k Da. It had high thermostability at 50°C and 60°C. At tested conditions, SOD was relatively stable in the presence of some inhibitors and denaturants, such as β-mercaptoethanol(β-ME), dithiothreitol(DTT), phenylmethylsulfonyl fluoride(PMSF), urea, and guanidine hydrochloride. Geobacillus sp. EPT3 SOD showed striking stability across a wide p H range from 5.0 to 11.0. It could withstand denaturants of extremely acidic and alkaline conditions, which makes it useful in the industrial applications.     

一个新的来自于弧菌QD-5并具有Poly-G内切功能的褐藻胶裂解酶的克隆和性质表征

Seven bacterial clones with alginate-utilizing activity were isolated from rotten kelp. By activity test, the Vibrio sp.QD-5 with the potential alginate-degrading capability was chosen to carry out the draft genome sequencing, and the result showed that the Vibrio sp. QD-5 containing an alginate lyase gene cluster. One of these genes, aly-IV,was cloned and characterized for the first time. After overexpression, Aly-IV, with a molecular mass of about62 kDa and a theoretical isoelectric point(pI) of 5.12, was purified to a specific activity of 1 256.78 U/mg and showed highest activity at 35°C in the Tris-HCl buffer at pH of 8.9. Moreover, the enzyme activity was enhanced by the metal ions of Na~+, K~+ and Mg~(2+) under certain concentration. Aly-IV degraded favorably polyG blocks in an endo-type, yielding monomer and dimer as the main products. Due to its high substrate specificity, Aly-IV could be used as a potential tool for production of polyG oligosaccharides with low degree of polymerization(DP) and for determining the fine structure of alginate.     

The influence of main ecological and environmental factors on the energy budget of Schlegel''s black rockfish Sebastes schlegeli

In the Maidao Ecological Laboratory of the Huanghai Sea Fisheries Research Institute in 1998 ~ 2000, the continuous flow-through method was adopted to determine the energy budget components and models of Schlegel＇ s black rockfish under different ecological and environmental factors, such as temperature, food species, body mass and feeding level, etc. The body mass of test fish ranged from 21.2 to 224.5 g. The specific growth rate tended to invert ＂U＂ change with temperature increase. The ecological conversion efficiency showed the same trend as temperature and feeding level rise. Only the specific growth rate tended to decelerate the increment with the rise of feeding level. From the above quantitative relationships, the maximum and optimal growth temper- atures could be obtained as 17.5 and 16.2 ℃, and the maintenance feeding rate and optimum feeding rate as 0.8% and 4. 1% of the body mass. The specific growth rate and ecological conversion coefficient showed a decelerating decrease trend with body mass increment. The small-sized fish food was more advantageous to the feeding level increase in comparison with the small-sized shrimp food, which led to the increase of growth, metabolism and excretion, but had no significant influence on the ecological conversion coefficient. All of temperature, feeding level, body mass and food species can obviously change the energy budget models. The assigning rate of growth energy tended to change along the invert ＂U＂ curve with the increase of temperature and feeding level, and decrease with the body mass increase, but the assigning rates of metabolism energy and excretion energy were quite reverse. These relationships could be quantitatively described as quadratic or power function curve. If the above relationships proved true to other marine fish, the establishment of energy budget model of the same fish under different conditions can be largely simplified.     

Changes of visceral properties and digestive enzymes in the herbivorous marine teleost Siganus canaliculatus fed on different diets

The rabbitfish Siganus canaliculatus is one of the few cultured herbivorous marine teleosts. To better understand the digestive physiology of this fish and provide data for designing formulated feed using macroalgae as an ingredient, the changes of visceral properties and digestive enzyme activities were investigated after the juveniles were fed on different types of food including raw fish(RF), formulated diet(FD) or macroalgae Enteromorpha prolifra(EP) and Gracilaria lemaneiformis(GL) for eight weeks. The results showed that the hepatosomatic and viscerosomatic indices in the RF and FD groups, as well as the relative intestine length(RIL) in the EP and GL groups, were significantly higher than those in other groups. Additionally, differences in the histological structure of the liver and anterior intestine were also observed among different dietary groups. The hepatic nuclei were displaced to the periphery by lipid inclusions in fish fed RF. The highest levels of mucosal folds were found in the anterior intestines of fish fed macroalgae. Digestive enzyme activity profiles showed obvious fluctuations in the first three weeks, and then leveled off in the following weeks. The levels of protease, lipase and α-amylase in the alimentary tract showed changes related to the levels of dietary protein, lipid and carbohydrate, respectively.Although macroalgae significantly inhibited the activity of protease in the stomach, it increased RIL and the number of mucosal folds in the anterior intestine so as to compensate for the influences on protease activities in the stomach. This study suggests that the digestive tract of rabbitfish can well adapt to different diets, and needs about three weeks to physiologically acclimatize to the nutritional status, thus implying that rabbitfish are somewhat omnivorous.     

大型水母沙海蜇暴发的温带贫营养海洋生态系统中细菌和浮游植物的关系

Bacterial abundance, phytoplankton community structure and environmental parameters were investigated to study the relationships between bacteria and phytoplankton during giant jellyfish Nemopilema nomurai blooms in the central Yellow Sea during 2013. N. nomurai appeared in June, increased in August, reached a peak and began to degrade in September 2013. Results showed that phosphate was possible a key nutrient for both phytoplankton and bacteria in June, but it changed to nitrate in August and September. Phytoplankton composition significantly changed that pico-phytoplankton relative biomass significantly increased, whereas other size phytoplankton significantly decreased during jellyfish bloom. In June, a significantly positive correlation was observed between chlorophyll a concentration and bacterial abundance(r=0.67, P0.001, n=34).During jellyfish outbreak in August, there was no significant correlation between phytoplankton and bacteria(r=0.11, P0.05, n=25), but the relationship(r=0.71, P0.001, n=31) was rebuilt with jellyfish degradation in September. In August, small size phytoplankton occupied the mixed layer in offshore stations, while bacteria almost distributed evenly in vertical. Chlorophyll a concentration significantly increased from(0.42±0.056) μg/L in June to(0.74±0.174) μg/L in August, while bacterial abundance just slightly increased. Additionally, the negative net community production indicated that community respiration was not entirely determined by the local primary productivity in August. These results indicated that jellyfish blooms potentially affect coupling of phytoplankton and bacteria in marine ecosystems.     

温度、盐度和光照对浒苔生长和光合作用的影响

Intensive Pyropia aquaculture in the coast of southwestern Yellow Sea and its subsequent waste, including disposed Ulva prolifera, was speculated to be one of the major sources for the large-scale green tide proceeding in the Yellow Sea since 2007. It was, however, unclear how the detached U. prolifera responded and resumed growing after they detached from its original habitat. In this study, we investigated the growth and photosynthetic response of the detached U. prolifera to various temperature, salinity and irradiance in the laboratory. The photosynthetic rate of the detached U. prolifera was significantly higher at moderate temperature levels(14–27°C)and high salinity(26–32), with optimum at 23°C and 32. Both low(14°C) and highest temperature(40°C), as well as low salinity(8) had adverse effects on the photosynthesis. Compared with the other Ulva species, U. prolifera showed higher saturated irradiance and no significant photoinhibition at high irradiance, indicating the great tolerance of U. prolifera to the high irradiance. The dense branch and complex structure of floating mats could help protect the thalli and reduce photoinhibition in field. Furthermore, temperature exerted a stronger influence on the growth rate of the detached U. prolifera compared to salinity. Overall, the high growth rate of this detached U. prolifera(10.6%–16.7% d~(–1)) at a wide range of temperature(5–32°C) and salinity(14–32) implied its blooming tendency with fluctuated salinity and temperature during floating. The environmental parameters in the southwestern Yellow Sea at the beginning of green tide were coincident with the optimal conditions for the detached U. prolifera.     

“海参-对虾”混养海水池塘的食物网结构及营养层级研究

The food sources of aquacultured Apostichopus japonicus and the trophic levels of organisms in a sea cucumber(A. japonicus) and prawn(Penaeus japonica) polyculture system in a saltwater pond in Zhuanghe, Liaoning Province were examined using carbon and nitrogen stable isotopes. Across organisms, δ13C ranged from(–25.47±0.20)‰ to(–16.48±0.17)‰(mean±SD), and δ15N ranged from(4.23±0.49)‰ to(12.44±0.09)‰. The δ13C and δ15N contents of A. japonicus, P. japonica and Fenneropenaeus chinensis were comparatively higher than those of other organisms. Values of δ13C and δ15N revealed that P. japonica, Hemigrapsus sanguineus and Neomysis japonica comprised the largest component of the diet of A. japonicus. The mean trophic level of the organisms in this saltwater pond polyculture system was(2.75±0.08). P. japonica, A. japonicus, F. chinensis,Synechogobius hasta and Neomysis japonica were in the 3rd trophic level(2–3); jellyfish, H. sanguineus and zooplankton were in the 2nd trophic level(1–2); and Enteromorpha prolifera, benthic microalgae, periphyton and suspended matter primarily consisting of phytoplankton, bacteria and humus were in the primary trophic level(0–1).     

Integration of the nuclease protection assay with sandwich hybridization (NPA-SH) for sensitive detection of Heterocapsa triquetra

Microalgae are photosynthetic microorganisms that function as primary producers in aquatic ecosystems. Some species of microalgae undergo rapid growth and cause harmful blooms in marine ecosystems. Heterocapsa triquetra is one of the most common bloom-forming species in estuarine and coastal waters worldwide. Although this species does not produce toxins, unlike some other Heterocapsa species, the high density of its blooms can cause significant ecological damage. We developed a H. triquetra species-specific nuclease protection assay sandwich hybridization (NPA-SH) probe that targets the large subunit of ribosomal RNA (LSU rRNA). We tested probe specificity and sensitivity with five other dinoflagellates that also cause red tides. Our assay detected H. triquetra at a concentration of 1.5×10⁴ cells/mL, more sensitive than required for a red-tide guidance warning by the Korea Ministry of Oceans and Fisheries in 2015 (3.0×10⁴ cells/mL). We also used the NPA-SH assay to monitor H. triquetra in the Tongyeong region of the southern sea area of Korea during 2014. This method could detect H. triquetra cells within 3 h. Our assay is useful for monitoring H. triquetra under field conditions.     

A new formula for saturated water steam pressure within the temperature range −25 to 220°C

Instead of approximation formula ln(E(t)/E(0)) = [(a ? bt)t/(c + T)] commonly used at present for representing dependence of pressure of saturated streams of liquid water E upon temperature we suggested new approximation formula of greater accuracy in the form ln(E(t)/E(0)) = [(A ? Bt + Ct ²)t/T], where t and T are temperature in °C and K respectively. For this formula with parameters A = 19.846, B = 8.97 × 10^?3, C = 1.248 × 10^?5 and E(0) = 6.1121 GPa with ITS-90 temperature scale and for temperature range from 0°C to 110°C relative difference of approximation applying six parameter formula by W. Wagner and A. Pruß 2002, developed for positive temperatures, is less than 0.005%, that is approximately 15 times less than accuracy obtained with the firs formula. Increase of temperature range results in relative difference increasing, but for even temperature range from 0°C to 220°C it does not higher than 0.1%. For negative temperatures relative difference between our formula and a formula of D. M. Murphy and T. Koop, 2005, is less than 0.1% for temperatures higher than ?25°C. This paper also presents values of coefficients for approximation of Goff and Grach formula recommended by IMO. The procedure of finding dew point T _d for known water steam pressure e _n based on our formula adds up to solving an algebraic equation of a third degree, which coefficients are presented in this paper. For simplifying this procedure this paper also includes approximation ratio applying a coefficient A noted above, in the form T _d (e _n ) = \(\frac{{AT_0 }}{{A - \varepsilon }}\) + 0.0866?² + 0.0116?^10/3, where ? = ln(e _n /E(T ₀)). Error of dew point recovery in this ratio is less than 0.005 K within the range from 0 to 50°C.     

Retrieval of total ozone in the mesosphere with a new model of electronic-vibrational kinetics of O<Subscript>3</Subscript> and O<Subscript>2</Subscript> photolysis products

The paper presents a new model of electronic-vibrational kinetics of the products of ozone and molecular oxygen photodissociation in the terrestrial middle atmosphere. The model includes 45 excited states of the oxygen molecules O₂(b ¹, Σ _g ⁺ ,v= 0−2), O₂ (a ¹Δ _g , v= 0−5), and O₂(X ³Σ _g ⁻ , v= 1−35) and of the metastable atom O (¹ D) and over 100 aeronomic reactions. The model takes into account the dependence of quantum yields of the production of O₂(a ¹Δ _g , v= 0−5) in a singlet channel of ozone photolysis in the Hartley band on the wavelength of photolytic emission. Taking account of the electronic-vibrational kinetics is important in retrieval of the vertical profiles of ozone concentration from measured intensities of the Atm and IR Atm emissions of the oxygen bands above 65 km and leads to an increase in the ozone concentration retrieved from the 1.27-μm emission, in contrast to the previous model of pure electronic kinetics. Sensitivity analysis of the new model is made for variations in the concentrations of atmospheric constituents ([O₂], [N₂], [O(³P)], [O₃], [CO₂]), the gas temperature, rate constants of the reactions, and quantum yields of the reaction products. A group of reactions that most strongly affect the uncertainty of ozone retrieval from measured intensities of atmospheric emissions of molecular oxygen O₂(b ¹Σ _g ⁺ , v) and O₂(a ¹Δ _g , v) has been determined. Original Russian Text ? V.A. Yankovsky, V.A. Kuleshov, R.O. Manuilova, A.O. Semenov, 2007, published in Izvestiya AN. Fizika Atmosfery i Okeana, 2007, Vol. 43, No. 4, pp. 557–569.     

基于稳定同位素分析推定秋冬季南极半岛南极大磷虾营养变化

南极大磷虾（Euphausia superba）为南大洋生态系统中的关键种,也是南极生态系统食物网中的重要枢纽。该种秋冬季转换期的营养信息对于理解其知之甚少的越冬机制非常重要。但关于此方面的少数研究在时空变化上仍存在着差异。为此,我们调查了南极半岛秋季（4-5月）和冬季（6月）磷虾成体δ¹³C和δ¹⁵N值的个体、月份及区域性差异。我们的目标旨在检验该期间磷虾的营养变化以及磷虾与其在南极海洋生态系统中的摄食环境之间的关系。结果如下：（1）磷虾δ¹³C值与体长之间无显著关系,但δ¹⁵N值与体长之间则存在显著相关性;（2）秋季磷虾δ¹³C值呈现增长趋势,但冬初季节并无显著变化,此期间δ¹⁵N值无显著不同;（3）布兰斯菲尔德与南设得兰群岛之间的δ¹⁵N平均值显著不同。我们的数据表明南极半岛秋至初冬转换期间磷虾成体营养呈现个体、季节性及区域性变化。     

太平洋褶柔鱼冬生群资源丰度变动对ENSO事件的响应研究

太平洋褶柔鱼为大洋性经济鱼种,具有一年生命周期,其资源变动受气候和海洋环境条件的显著影响。本研究根据日本提供的2003-2012年太平洋褶柔鱼冬生群体的渔业统计数据,结合产卵场环境数据以及尼诺指数ONI（定义为Niño 3.4区海表温度距平值）,分析不同气候条件下（厄尔尼诺和拉尼娜）太平洋褶柔鱼冬生群体产卵场海表温度（SST）、叶绿素a（Chl-a）浓度以及适宜产卵面积（SSA）的变动情况及对其资源丰度（CPUE）的影响。结果表明,太平洋褶柔鱼冬生群体产卵场SST、Chl-a浓度和SSA具有明显的季节性变化。相关分析表明,各年CPUE与Chl-a浓度以及SSA具有显著的正相关关系（p<0.05）,但与SST相关性不显著（p>0.05）。此外,厄尔尼诺和拉尼娜事件通过驱动太平洋褶柔鱼冬生群体产卵场SSA和关键海域（25°-29°N,122.5°-130.5°E）内的Chl-a空间分布和大小变化,从而改变其资源丰度,但影响作用随各异常事件的强度不同而变化,具体表现为：发生弱强度厄尔尼诺事件时,产卵场SSA较高,Chl-a浓度处于较低水平,导致资源补充量处于较低水平,CPUE降低;发生中等强度厄尔尼诺事件时,产卵场SSA较低,但Chl-a浓度处于较高水平,导致资源补充量增加,CPUE处于上升水平;发生中等强度拉尼娜事件时,产卵场SSA和Chl-a浓度均处于较高水平,资源补充量显著增加,CPUE显著升高。研究表明,厄尔尼诺和拉尼娜事件对太平洋褶柔鱼冬生群体产卵场摄食孵化环境和资源丰度变动具有显著影响。