A multi-generation Schmakeria poplesia culturing system for use in ecotoxicological study

Crustacean zooplankton form the keystone link between primary producers and fish stocks in marine and estuary ecosystems. We have established a multi-generation cultivation system for zooplankton with which future experiments on the biological effects of pollutants in marine and estuary environments can be better performed. A population of calanoid copepod, Schmakeria poplesia, was collected in December 2003 and maintained in a static system through all stages (eggs to adults). The population exhibited an average developmental time of 13.6 d in conditions corresponding to the natural environment (water temperature 20°C, salinity 15). A series of experiments were performed to examine copepod egg production and hatching success as functions of food type and feeding concentration. Results in our study showed that Isochrysis galbana was more favored for the reproduction of copepods than Phaeodactylum tricornutum, and 10×10⁴cells mL⁻¹ was the most practical algae concentration. We have demonstrated that the Schmakeria poplesia population can be maintained in the laboratory through multiple generations. In addition, methods to control egg production through changes in food concentration have been established, making it feasible to control the start date of exposure experiments or the timing of the collection of offspring to initiate a new generation.     

Modeling the Roles of Precipitation Increasing in Glacier Systems Responding to Climate Warming —— Taking Xinjiang Glaciated Region as Example

The studies on prediction of climate in Xinjiang almost show that the precipitation would increase in the coming 50 years, although there were surely some uncertainties in precipitation predictions. On the basis of the structure of glacier system and nature of equilibrium line altitude at steady state （ELAo）, a functional model of the glacier system responding to climate changes was established, and it simultaneously involved the rising of summer mean temperature and increasing of mean precipitation. The results from the functional model under the climatic scenarios with temperature increasing rates of 0.01, 0.03 and 0.05 K/year indicated that the precipitation increasing would play an evident role in glacier system responding to climate change： if temperature become 1 ℃ higher, the precipitation would be increased by 10%, which can slow down the glaciers retreating rate in the area by 4 %, accelerate runoff increasing rate by 8 % and depress the ELAo rising gradient by 24 m in northern Xinjiang glacier system where semi-continental glaciers dominate, while it has corresponding values of only 1%, 5 % and 18m respectively in southern Xinjiang glacier system, where extremely continental glaciers dominate.     

Bacteriological analysis of the digestive tube of the mud snail (Bullacta exarata Philippi) and its rearing shoal

The bacterial flora in the digestive tube of Bullacta exarata Philippi and its rearing shoal were investigated. A total of 157 strains of heterotrophic bacteria, isolated from crop, stomach intestine and other parts of the digestive tube, mainly belong to the genera Photobacterium, Bacillus, Pseudomonas Vibrio and some genera of the family Enterobacteriaceae. There are significantly more varieties of bacteria in crop than in stomach and intestine. A total of 173 strains of bacteria were isolated from the rearing shoal, belonging to 13 genera. The 5 predominant genera, such as Bacillus and Photobacterium, are the same as those in the digestive tube, but greatly differ in percentages. The number of heterotrophic bacteria and Vibrio in rearing shoal change in line with the alteration of the temperature, and are significantly affected by the use of pesticides.     

Seasonal change in body length of important small copepods and relationship with environmental factors in Jiaozhou Bay,China

Differences among species in prosome length and in species’ response to environmental factors do exist. Therefore, it is useful to examine prosome length for different copepod species in variable environments. Seasonal variations in prosome length of four small copepods and their copepodite stages in the Jiaozhou Bay were compared and the relative influence of temperature, salinity, and chlorophyll concentration were examined. Two peaks were found in the mean prosome length of Paracalanus parvus (in early winter and May). For Acartia bifilosa, the maximum values of all copepodites occurred mainly from February to April, and decreased to the bottom in July. Prosome length of Acartia pacifica peaked when it first appeared in June, then reached to the minimum in July. Parvocalanus crassirostris only appeared from late summer to autumn and the mean prosome length showed no clear changes. Correlations of adult prosome length with environmental factors were evaluated. For the four species, temperature was negatively correlated to prosome length except for P. crassirostris. But the different species varied markedly in their responds to temperature. A. bifilosa showed a more definite trend of size variation with temperature than P. parvus and A. pacifica. Correlations of prosome length with salinity were significantly positive for almost all the small copepods. The relationship between chlorophyll concentration and prosome length was complicated for these copepods, but for P. parvus, chlorophyll concentration was also an important affecting factor. Furthermore, investigation needs to be done on food quality for some copepod. These results are essential to estimate the biomass and the production, and to understand these small copepods’ population dynamics in this human-affected bay.     

Cohesive strength and seismogenic stress pattern along the active basement faults of the Precordillera-Sierras Pampeanas ranges, western Argentina: An experimental analysis by means of numerical model

A two-dimensional finite element method （FEM） model that incorporates faults, elastic rock physical properties, topographical load due to gravity and far-field plate velocity boundary conditions was used to recognize the seismogenic stress state along the fold-and-thrust belt of the Precordillera-Sierras Pampeanas ranges of western Argentina. A plane strain model with nine experiments was presented here to examine the fault strength with two major rock phyical properties： cohesion and angle of internal friction. Mohr-Coulomb failure criterion with bulk rock properties were applied to analyse faults. The stress field at any point of the model was assumed to be comprised of gravitational and tectonic components. The analysis was focused to recognize the seismogenic shear strain concentrated in the internal-cristaline domain of the orogene shown by the modeling. Modeling results are presented in terms of four parameters, i. e., （i） distributions, orientations, and magnitudes of principal stresses （σ1 and σ3）, （ii） displacement vector1 （iii） strain distribution, and （iv） maximum shear stress （τmax） contour line within the model. The simulation results show that the compressive stress is distributed in and around the fault systems. The overall orientation of of σ1 is in horizontal directions, although reorientations do occur within some stress weaker parts, especially subsequent to the faults. A large-scale shear stress is accumulating along the active faults of Tapias-Villicum Fault （TVF）, Salinas-Berros Fault （SBF）, Ampacama-Niquizanga Fault （ANF） and Las Charas Fault （CF）, which could act as local stress and strain modulators to localize the earthquakesoccurrence.     

Molecular identification and culture observation on <Emphasis Type="Italic">Acrochaete leptochaete</Emphasis> (Chaetophoraceae,Chlorophyta) from China

Acrochaete leptochaete (Huber) Nielsen (Chaetophoraceae, Chlorophyta) was isolated from the macroalgae Chaetomorpha collected from intertidal pools in Rongcheng, Shandong, China. 18S rDNA combined with ITS regions were used to ascertain the morphological identification of the isolated material. Based on the unialgal culture, asexual reproduction and growth characteristics of A. leptochaete were investigated over wide ranges of temperature and irradiance. Results revealed that asexual reproduction of A. leptochaete could be realized by biflagellate zoospores. The zoospores germinated directly to give self- replicating generations. Zoospore germination was bipolar. A temperature range from 13-21°C and a lower irradiance of 36 μmol/(m2·s) were most favorable for the growth of A. leptochaete. Thallus organization, an important taxonomic criterion for the genus Acrochaete, was affected markedly by temperature and irradiance. Our results extend the knowledge about the species’ general biology and its morphological plasticity. For classification and identification of a simple microphytic algae like A. leptochaete, which are traditionally placed in the class Chaetophoraceae, we propose that molecular tools associated with culture observations are applied.     

Characterization of Pythium chondricola associated with red rot disease of Pyropia yezoensis (Ueda) (Bangiales,Rhodophyta) from Lianyungang,China

Pyropia yezoensis(formerly Porphyra yezoensis) is an economically important red alga that is cultured extensively in China. The red rot disease occurs commonly during Pyropia cultivation,causing serious economic losses. An incidence of red rot disease was found in a P. yezoensis farm from mid-November to mid-December 2015 at Lianyungang, Jiangsu Province, China. Histopathological examination revealed that the naturally infected thalli were infected apparently by a pathogen, leading to red rot symptoms. The causative agent was isolated and identified as the oomycete Pythium chondricola by morphological analysis and sequence analysis of the internal transcribed spacer and cytochrome oxidase subunit 1(cox1). In artificial infection experiments on the P. yezoensis blades, the P. chondricola isolate was able to cause the same characteristic histopathology seen in natural infections. P. chondricola grew well at a wide range of temperatures in the range 8-31℃, salinities at 0-45 and pH 5-9. In an orthogonal test used to determine the effects of environmental factors(temperature, salinity, and zoospore concentration)on infection, the data revealed that temperature was the most important factor to affect red rot disease development, with the optimal conditions for disease expansion being 20℃, 35 salinity, and a zoospore concentration of 10~6 zoospores/mL. The results obtained from the present study prompted us to set up a comprehensive epidemiological study on Pyropia, which will provide support to maintain the healthy development of the Pyropia industry in China.     

Establishment and characterization of a new cell line from the kidney of spotted halibut <Emphasis Type="Italic">Verasper variegates</Emphasis>

A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subcultured more than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ml-1 basic fibroblast growth factor(bFGF).Cell morphology from primary culture and subculture was observed continuously by microscopy.The SHK cell line consisted predominantly of fibroblast-like cells.The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃.The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%.The doubling time of the cells was determined to be 44.8 h.Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number (2n=46).The cells were successfully transfected with green fluorescent protein(GFP)reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies.The cells were infected by lymphosystis disease virus(LCDV)and found to be susceptible to the virus in cytopathic effect(CPE)observation.The infection was confirmed by PCR and electron microscopy experiments,which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.     

PROFESSOR ZHU KEZHEN OPENING UP A PATH FOR RESEARCH ON CLIMATIC CHANGE IN CHINA

ＰＲＯＦＥＳＳＯＲ ＺＨＵ ＫＥＺＨＥＮ ＯＰＥＮＩＮＧ ＵＰ Ａ ＰＡＴＨ ＦＯＲ ＲＥＳＥＡＲＣＨ ＯＮ ＣＬＩＭＡＴＩＣ ＣＨＡＮＧＥ ＩＮ ＣＨＩＮＡ ＳｈｉＹａｆｅｎｇ（施雅风）（ＬａｎｚｈｏｕＩｎｓｔｉｔｕｔｅｏｆＧｌａｃｉｏｌｏｇｙａｎ...     

Establishment of a spleen cell line from large yellow croaker Pseudosciaena crocea and its primitive application in foreign gene transfection

A large yellow croaker,Pseudosciaena crocea,spleen(LYCS)cell line was established and the feasibility of using it for foreign gene transfection was evaluaed in this study.Primary culture of LYCS cells was initiated from spleen tissue pieces,which were cultured at 25℃ in Dulbecco’s modiced Eagle medium/F12 medium(DMEM/F12,1:1)(pH7.2),supplemented with 20% fetal bovine serum,carboxymethyl chitosan,chondroitin sulfate,basic fibroblast growth factor(bFGF)and insulin-like growth factor-I(IGF-I).The cultured LYCS cells,in fibroblast shape,proliferated to 100% confluency 20 days later.Chromosome analyses indicated that the LYCS cells exhibited chromosomal aneuploidy with a modal chromosome number of 48 which displayed the normal diploid karyotype of P.crocea(6m+6sm+36t,NF=60).A LYCS cell line,with a population doubling time of 48.7 h at passage 60,has been established and subcultured to passage 70.Transgenic feasibility test demonstrated that positive green fluorescence protein(GFP)expression was observed in LYCS cells after pcDNA3.1-GFP plasmid transfection.In conclusion,a continuous foreign gene trans-fection feasible LYCS cell line has been established successfully.The cell line might serve as a valuable tool for studies of transgenic breeding and has potential applications for different kinds of cytotechnological studies.     

An extracellular oligopeptide permease may be a potential virulence factor of Vibrio harveyi

An oligopeptide permease A(OppA)was purified from the extracellular product of Vibrio harveyi SF-1.The molecular weight of the purified protein was estimated to be 58 kDa on SDS-PAGE.The purified protein showed phospholipase C activity at the optimal values of temperature 50℃ and pH 8.0.The enzymatic activity decreased when the temperature increased to 40℃.The N-terminal sequence of the purified protein was determined as ADVPAGTKLA,which is similar to that of OppA.The OppA pre-cursor gene was cloned from th...     

Induction and cultivation of cloned filaments of Polysiphonia urceolata (Rhodomelaceae, Rhodophyta)

A filamentous clone of Polysiphonia urceolata was regenerated from segments cut from the fronds of gametophytes. Unlike wild thalli with short virgate branchlets, the clone was filamentous with few branches. Many transparent trichoblasts arose from pericentral cells during the induction culture, but these were seldom observed during normal growth. The trichoblasts were uniseriate, often colorless, and formed lobed rhizoids rapidly when they came into contact with solid substrates. In addition to morphological characteristics, the photosynthetic properties and growth conditions of the clone differed from those of the mother plant. Cross-gradient light and temperature culture experiments revealed that the most favorable conditions for culture of the filamentous clone were 22°C and 95-120 μE/(m2-s) light intensity. The photosynthetic light saturation value for filaments was approx. 100 μE/(m2-s), which is far lower than that of wild thalli. These results could be used to develop techniques for mass cultures of P. urceolata in photobioreactors for production of seed stock or bioactive products.     

The effects of temperature and substrate on ontogenetic behavior of bastard halibut, Paralichthys olivaceus （T.et S.）

The effects of temperature and substrate on ontogenetic patterns and settlement behavior of bastard halibut, Paralichthys olivaceus （T. et S.） were studied in a series of laboratory experiments. Analysis on stage-based data revealed that except for settlement, and compared to the substrate, the ambient temperature dominantly controlled the hatch success and subsequent development of the species. The oosperm optimum survival rate of 60% during hatching occurred at 20℃, and survival rate of 29%, at 18℃ for larval rearing. The survival rate of larvae reared under variable temperature of 3.5-4.5℃ was higher than that of those reared under stable temperature. The maximum survival rate of 29%, corresponded to variation at temperature of 4℃. With increase in age newly settled juveniles preferred to lie on the fine substrate and gradually moved on the coarse substrate. GLM （general linear model） analysis showed that the combination of temperature and substrate had no significant impact on the survival of settling larvae, but definitely affected the duration of settlement.     

Relationship between heavy metal pollution and water productivity in Xiamen estuarine harbor

This paper is based on the results of simulation experiments and annual observations on the effect of Cr, Cu, Hg, and Zn on diatom (Chaetoceros sp.Skeletonema costatum andMelosira sulcata) growth. Laboratory experiments reveal many physico-chemical parameters play an important role in the toxicity and accumulation of metals in organisms. The mesotrophic level in the water environment is related to phytoplankton growth. A higher nutrient level can have a negative impact on diatom production. Several of the above metals coexist in the investigated region and exert mainly a negative effect on growth. This paper was published in Chinese in theOceanologia et Limnologia Sinica 17(3):173–183, 1986.     

The effect of temperature on growth and energy budget of the polychaete, Neanthes japonica Izuka

Growth and energy budget of the polychaete, Neanthes japonica, at various temperatures (17, 20, 23, 26 and 29°) were investigated in this study. The growth, as indicated by final dry weight and specific growth rate (SGR), increased with increasing temperature, with the maximum level at 26°C, and then decreased significantly at 29°C. A similar trend was observed in feeding rate, food conversion efficiency (FCE) and apparent digestive rate (ADR). However, no significant differences were detected in ADR among all the temperature treatments. In the pattern of energy allocation, faeces energy was only a small component of energy budget and had little influence on the proportion of food energy allocated to growth. The metabolic energy accounted for a large portion of energy intake for each temperature treatment. The nitrogen excretion was appreciable with changing temperature. The two expenditure terms (respiration energy and excretion energy) in energy budget were the major factors influencing the proportion of food energy allocated to growth. These results revealed that temperature affected the growth of N. japonica mainly by influencing feeding rate and FCE. In addition, regression equations describing the relationship between feeding rate, faecal production, SGR, FCE and temperature were obtained. The optimum temperatures for feeding rate, FCE and SGR were estimated at 25.01°C, 24.24°C and 24.73°C, respectively, from the regression equations.     

Simultaneous estimates ofsynechococcus spp. Growth and grazing mortality rates in the English Channel

The marine chroococooid phycoerythrin-containingSynechococcus spp. cyanobacterium has been implicated as a substantial component of the photosynthetic picoplankton in the ocean. Although its importance as food source for heterotrophic nanoplankton is now recognized, information about the cycling ofSynechococcus biomass and its diel pattern is limited and study methodology varies among authors. The selective metabolic inhibitor method was used to simultaneously estimate growth and grazing disappearance rates ofSynechococcus in the English Channel where growth rates ranged from 0.25 to 0.72/d (mean ±SD=0.51±0.17/d) and grazing mortality rates ranged from 0.19 to 0.64/d (mean ±SD=0.48±0.17/d). Size-fractionated experiments demonstrated that up to 70% ofSynechococcus disappearance could be attributed to grazers going through a 2 μm Nuclepore filter.Synechococcus grazing mortality rates (mean=0.74 ±0.25/d) during the day were always higher than that (mean=0.2±0.20/d) during the night, while growth rates showed no clear diel pattern. A positive correlation was observed between growth rates andin situ temperature ranging from 9 to 17°C, while in contrast grazing was independent of temperature. The close similatiry between average growth and grazing rates suggests a rapid recycling ofSynechococcus biomass in English Channel coastal waters.     

Impacts of Climatic Factors on Runoff Coefficients in Source Regions of the Huanghe River

Runoff coefficients of the source regions of the Huanghe River in 1956–2000 were analyzed in this paper. In the 1990s runoff of Tangnaihai Hydrologic Station of the Huanghe River experienced a serious decrease, which had at- tracted considerable attention. Climate changes have important impact on the water resources availability. From the view of water cycling, runoff coefficients are important indexes of water resources in a particular catchment. Kalinin baseflow separation technique was improved based on the characteristics of precipitation and streamflow. After the separation of runoff coefficient (R/P), baseflow coefficient (Br/P) and direct runoff coefficient (Dr/P) were estimated. Statistic analyses were applied to assessing the impact of precipitation and temperature on runoff coefficients (including Dr/P, Br/P and R/P). The results show that in the source regions of the Huanghe River, mean annual baseflow coefficient was higher than mean annual direct runoff coefficient. Annual runoff coefficients were in direct proportion to annual pre- cipitation and in inverse proportion to annual mean temperature. The decrease of runoff coefficients in the 1990s was closely related to the decrease in precipitation and increase in temperature in the same period. Over different sub-basins of the source regions of the Huanghe River, runoff coefficients responded differently to precipitation and temperature. In the area above Jimai Hydrologic Station where annual mean temperature is –3.9oC, temperature is the main factor in- fluencing the runoff coefficients. Runoff coefficients were in inverse relation to temperature, and precipitation had nearly no impact on runoff coefficients. In subbasin between Jimai and Maqu Hydrologic Station Dr/P was mainly affected by precipitation while R/P and Br/P were both significantly influenced by precipitation and temperature. In the area be-tween Maqu and Tangnaihai hydrologic stations all the three runoff coefficients increased with the rising of annual precipitation, while direct runoff coefficient was inversely proportional to temperature. In the source regions of the Huanghe River with the increase of average annual temperature, the impacts of temperature on runoff coefficients be-come insignificant.     

The application of fractal study in forecasting flood calamities in Wuzhou, Guangxi

１ＦＲＡＣＴＡＬＴＨＥＯＲＹＡＮＤＦＬＯＯＤＣＡＬＡＭＩＴＩＥＳＴｈｅｍｏｓｔｒｅｇｕｌａｒｇｅｏｍｅｔｒｉｃｏｂｊｅｃｔｓｗｈｉｃｈｃｏｕｌｄｂｅｍｅａｓｕｒｅｄｂｙｌｅｎｇｔｈｓｏｆｓｐｅｃｉｆｉｃｉｔｙａｒｅｔｈｅｉｓｓｕｅｏｆｃｌａｓｉｃａ...     

Simulation and analysis on seasonal variability of average salinityin the Yellow Sea

The CTD (conductivity, temperature and depth) data collected by six China-Korea joint cruises during 1996-1998 and the climatological data suggest that the seasonal variability of average salinity in the Yellow Sea (Sa) presents a general sinusoid pattern. To study the mechanism of the variability, annual cycles of Sa were simulated and a theoretical analysis based on the governing equations was reported.Three main factors are responsible for the variability: the Yellow Sea Warm Current (YSWC), the Changji-ang (Yangtze) River diluted water (YRDW) and the evaporation minus precipitation (E-P). From December to the next May, the variability of Sa is mainly controlled by the salt transportation of the YSWC. But in early July, the YSWC is overtaken and replaced by the YRDW which then becomes the most important controller in summer. From late September to November, the E-P gradually took the lead. The mass exchange north of the 37癗 line is not significant.     

Construction of Porphyra yezoensis Pure Line from Protoplasts and Its 18S rDNA Sequence Determination

The wild Porphyra yezoensis collected from the Qingdao coast was used to prepare protoplasts by enzyme digestion. The pure line was constructed by cultivating the protoplasts. The 18S rDNA of the P. yezoensis pure line was cloned and sequenced. Sequence analysis was executed for this sequence and other 22 sequences retrieved from GenBank. A phylogenetic tree was constructed using the neighbor-joining method. The results revealed a high diversity of 18S rDNA sequences in genus Porphyra and the considerable variation of 18S rDNA sequences in different strains of the same species P. yezoensis and P. tenera. Significant difference of 18S rDNA sequence was observed between P. yezoensis from Qingdao, China, and the two strains of P. yezoensis from Japan, but the three strains of P. yezoensis formed a stable clade in the phylogenetic tree. These results indicate the possibility of interspecies and intraspecies discrimination of Porphyra using the 18S rDNA sequences.