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Starvation effects on pathogenic Vibrio alginolyticus in natural seawater

Acta Oceanologica Sinica 2008年第4期27卷 120-129页

作者： YI Jiabo CHEN Qiang ZOU Wenzheng YAN Qingpi ZHUANG Zhixia WANG Xiaoru Fisheries College Key Laboratory of Science and Technology for Aquaculture and Food Safety Jimei University Xiamen 361021 China First Institute of Oceanography State Oceanic Administration Qingdao 266061 China

To get a better understanding of the starvation survival strategy of pathogenic Vibrio alginolyticus, log-phase cells were inoculated into sterile natural seawater for starvation studies. The results showed that all of total bacteria number, viable bacteria number and CFU number of V. alginolyticus increased remarkably at the initial starvation stage; after reaching their peaks at 5 d, both total bacteria number and viable bacteria number of V. alginolyticus fell slowly, while the CFU number fell more quickly after reaching its peak at 10 d; V. alginolyticus elongated their cells at the prophase of starvation, and then shrunk their volume and turned their shapes into ovals from rods at the anaphase of starvation; starved cells showed more sensitivity to heating and UV; starved cells showed no significant difference from unstarved ones at the lowest detection limit determined by indirect enzyme-linked immu- nosorbent assay （ELISA） ; starved cells＇ ability to adhere to the skin mucus of large yellow croaker （ Pseudosciaena crocea） showed a sharp decline as the starvation time increases; the cellular protein of V. alginolyticus increased remarkably at the ariaphase of starvation. The results indicated that pathogenic V. alginolyticus could survive in starvation for relatively long periods of time （ ≥2 months） in 28℃ natural seawater due to the morphological and physiological changes; however, starved V. alginolyticus cells showed less virulence and higher sensitivity under environmental stresses.

关键词： Vibrio alginolyticus starvation survival adhesion ELISA resistance

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Adhesion of pathogenic Vibrio alginolyticus to the gill mucus of Pseudosciaena crocea

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Acta Oceanologica Sinica 2007年第3期26卷 101-109页

作者： CHEN Qiang YAN Qingpi MA Shen ZHUANG Zhixia WANG Xiaoru Fisheries College Jimei University Xiamen 361021 China Fisheries College Ocean University of China Qingdao 266003 China First Institute of Oceanography State Oceanic Administration Qingdao 266061 China

Adhesion of Vibrio alginolyticus to the gill mucus of Pseudosciaena crocea has been investigated using [ methyl-^3 H ] thymidine as isotope tracer. The results showed that： the adhesive quantity of V. alginolyticus increased with bacterial concentrations and reached equilibrium after incubated for 180 min; the higher adhesive quantity was obtained at 15 ~ 30 ℃ and sourish conditions; adhesion of V. alginolyticus could not achieved without Na^＋ , and Ca^2＋ played an auxiliary role in the bacterial adhesion; adhesion of V. alginolyticus was inhibited remarkably by starvation, heat treatment and periodic acid treatment; all of the eight kinds of carbohydrates investigated enhanced the adhesion of V. alginolyticus to the gill mucus of P. crocea, among them, glucose, mannose, fructose and maltose showed the specially enhanced adhesion. The results indicated that E alginolyticus could adhere to the gill mucus of P. crocea facilely in seawater, and this bacterial adhesion was influenced by environmental factors and closely related to superficial carbohydrate structures and some heat-sensitive structures.

关键词： Vibrio alginolyticus adhesion Pseudosciaena crocea gill mucus

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Non-specific immune response of bullfrog Rana catesbeiana to intraperitoneal injection of bacterium Aeromonas hydrophila

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Chinese Journal of Oceanology and Limnology 2008年第3期26卷 248-255页

作者：张俊杰邹文政鄢庆枇 Fisheries College Jimei UniversityXiamen 361021 China Chemistry and Chemical Engineering College of Xiamen University Xiamen 361005 China

Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two *** bullfrog in bacterium-injected group was injected intraperitoneally(i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 106 CFU/ml,while each one in control group injected *** 0.2 ml sterile saline solution(0.85%,w/v).Three bullfrogs in both groups were sampled at 0,1,3,7,11,15 and 20 days post-injection(dpi) for the evaluation of non-specific immune *** was observed that intraperitoneal injection of *** significantly increased the number of leucocytes and that of NbT-positive cells in peripheral *** increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control ***,a significant reduction was detected in vitro in phagocytosis activity of peripheral blood *** significant difference in changes in the number of peripheral erythrocytes,serum superoxide dismutase(SOD) activity,and lysozyme activity was detected between the two *** is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the *** infection.

关键词： Rana catesbeiana Aeromonas hydrophila intraperitoneal injection non-specific immune response

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The experimental studies on the carbon and nitrogen budgets of Pseudeuphausia sinica

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Acta Oceanologica Sinica 2003年第2期22卷 265-272页

作者： Guo Donghui, Li Shaqjing, Chen Feng, Wang Guizhong, Chen Gang Department of Oceanography,Institute of Subtropic Ocean,Xiamen University,Xiamen 361005,China Department of Oceanography Institute of Subtropic Ocean Xiamen University Xiamen 361005 China

The carbon and nitrogen budgets were estimated on the adult females, juveniles and post-furcilia larvae of Pseudeuphausia sinica fed on newly hatching nauplii of Artemia salina in the laboratory. It was found that the ingestion rate was linearly related to the food concentration, suggesting high feeding potential. The linear correlation could be established between the respirating rate (carbon consumption rate) and carbon ingestion rate, as well as carbon assimilation rate. The regression coefficients (*** dynamic action coefficients) were in the range from 9% to 16% (ingested C) or 10% to 17% (assimilated C) respectively, with lower in the post-furcilia larvae. There also existed a linear correlation equation between estimated total nitrogen excretion rate and the rates of nitrogen ingestion and assimilation separately, except for the juveniles. The defecation rates increased with the increase of the ingestion rate; as a result, assimilation efficiency was not related to the ingestion rate, ranging from 0.84 to 0.95. The results inducated that the nitrogen content in food particles was a key factor limiting the growth of P. sinica. The critical ingestion rate was 10 μgN·mg-1 body dry weight per day. Assimilated N was lost mostly by excretion, following allocated to somatic growth. The nitrogen loss by moult only accounted for a minor part. As for carbon budget, respiration and somatic growth also accounted for most of assimilation, but varied with ingestion rates. Moult loss was minor. Estimated reproductive growth (C&N) in the adult females accounted for somewhat higher percent of assimilation than the moult growth. The net growth efficiency (K2) increased with the increase of the ingestion rates, but decreased slightly for juvenile and post-furcilia larvae after the rates up to a certain value.

关键词： Pseudeuphausia. sinica carbon and nitrogen budgets Xiamen harbour

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Overexpression and characterization of a thermostable β-agarase producing neoagarotetraose from a marine isolate Microbulbifer ***1

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Acta Oceanologica Sinica 2019年第2期38卷 96-106页

作者： Yanbing Zhu He Gao Hebin Li Hui Ni Zedong Jiang Lijun Li Anfeng Xiao College of Food and biological Engineering Jimei University fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering Research Center of Food biotechnology of Xiamen City Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed Xiamen Southern Ocean Technology Center of China Department of Pharmacy Xiamen Medical College

An agarase gene containing 1 302 bp was cloned from Microbulbifer sp. AG1. It encoded a mature protein of 413 amino acids plus a 20-residue signal peptide. The recombinant enzyme without the signal peptide was expressed and purified from Escherichia coli bL21(DE3). When agarose was used as a substrate, the optimal temperature and pH for the enzyme were 60℃ and 7.5, respectively. The recombinant agarase showed excellent thermostability with 67% and 19% of residual activities after incubation at 50℃ and 60℃ for 1 h, *** SDS, the recombinant agarase had a relatively good resistance against the detected inhibitors, detergents and urea denaturant. Thin layer chromatography analysis and enzyme assay using p-nitrophenyl-α/β-Dgalactopyranoside revealed that the recombinant agarase was a β-agarase that degraded agarose into neoagarotetraose as the main end product. The enzymatic hydrolysis products with different degree of polymerization exhibited the antioxidant activities.

关键词： thermostable β-agarase neoagarotetraose Microbulbifer sp.

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Population structure of Haliotis rubra from South Australia inferred from nuclear and mtDNA analyses

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Acta Oceanologica Sinica 2006年第4期25卷 99-112页

作者： LI Zhongbao Sharon A Appleyard Nicholas G Elliott Fisheries College Jimei University Xiamen 361021 China Commonwealth Scientific and Industrial Research Organization Marine and Atmospheric Research GPO Box 1538 HobartTasmania 7001 Australia

Microsatellite loci and mtDNA-RFLPs were surveyed in four spatially separated populations of Haliotis rubra and two populations of putative Haliotis conicorpora from South Australia. A high level of microsatellite genetic diversity was observed in all populations although several loci were characterized by homozygote excesses, probably due to null alleles. MtDNA variation was also moderate with an average of 80% haplotype variation across the six populations. Despite the high levels of genetic variation, significant pair-wise spatial differentiation is not detected among the populations. Hierarchical AMOVA analysis revealed very low levels of genetic partitioning on either a spatial or putative species level. both molecular techniques revealed little genetic differentiation across the six populations, suggesting a panmictic population model for these South Australian abalone populations. Furthermore, no molecular evidence suggests that the putative H. conicorpora individuals sampled from South Australia belong to a separate species.

关键词： Haliotis rubra H. conicorpora South Australia genetic diversity genetic differentiation mtDNA-RFLPs microsatellites

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Substitution of His260 residue alters the thermostability of Pseudoalteromonas carrageenovora arylsulfatase

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Acta Oceanologica Sinica 2019年第6期38卷 75-82页

作者： Yanbing Zhu Xiaoqian Yin Han Liu Hebin Li Yanhong Chen Lijun Li Anfeng Xiao Hui Ni College of Food and biological Engineering Jimei UniversityXiamen 361021China fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering Xiamen 361021China Research Center of Food biotechnology of Xiamen City Xiamen 361021China Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed Xiamen Southern Ocean Technology Center of ChinaXiamen 361021China Department of Pharmacy Xiamen Medical CollegeXiamen 361023China

This study aimed to improve the thermostability of arylsulfatase from Pseudoalteromonas carrageenovora. A library of P. carrageenovora arylsulfatase mutants was constructed by introducing random mutagenesis using error-prone PCR. After screening, two mutants of H260L and D84A/H260L showed enhanced thermal stability than the wild-type predecessor (WT). Site-directed mutagenesis demonstrated that only amino acid residue at Position 260 plays an important role in the thermostability of P. carrageenovora arylsulfatase. Thermal inactivation analysis showed that the half-life (t1/2) values at 55°C for H260L, H260I, H260Q, H260F and H260R were 40.6, 48.4, 30.9, 29.1 and 34.5 min, respectively, while that of WT was 9.1 min. Structure modeling demonstrated that the additional hydrogen bonds and/or optimization of surface charge-charge interactions could be responsible for the increased thermostability imparted by H260L, H260I, H260Q, H260F and H260R.

关键词： arylsulfatase Pseudoalteromonas carrageenovora directed evolution error-prone PCR thermostability

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Discovery of Cladonema multiramosum ***.(Cnidaria:Hydrozoa:Cladonematidae)using DNA barcoding and life cycle analyses

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Acta Oceanologica Sinica 2022年第3期41卷 44-52页

作者： Konglin Zhou Yuping Gu Lu Wang Jianming Chen Institute of Oceanography College of Geography and OceanographyMinjiang UniversityFuzhou 350108China

In contrast to typical planktonic hydromedusae,Cladonema medusae are mostly benthic,with specialised adhesive branches to adhere to the *** this study,a Cladonema species discovered in a laboratory aquarium in Fuzhou,China,was confirmed as a new species,based on morphological and molecular *** species was named Cladonema multiramosum *** medusa is distinct from that of congeners possessing substantially more adhesive branches(8-24,rarely 5-7),and tiny branches on the upper radial *** validity of *** *** also supported by molecular phylogenetic analyses based on the mitochondrial 16S rDNA ***,*** *** also displayed considerable phenotypic plasticity with respect to its radial canals,tentacles,stinging branches per tentacle,oral tentacles,manubrium,and gonads,hindering species identification based solely on *** some morphological characteristics of hydroids(filiform tentacles and medusa buds)and nematocysts could also be used as diagnostic characters in the genus Cladonema,this information is unavailable for some Cladonema ***,the taxonomy within the genus Cladonema was re-evaluated based mainly on the morphological characteristics of the *** revision of the genus Cladonema should be made when supplementary information on the life cycle and DNA barcoding are updated.

关键词： Cladonema morphology life cycle DNA barcoding 16S rDNA

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Air-sea carbon fluxes and their controlling factors in the Prydz bay in the Antarctic

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Acta Oceanologica Sinica 2008年第3期27卷 136-146页

作者： GAO Zhongyong CHEN Liqi GAO Yuan Key Laboratory of Global Change and Marine Atmospheric Chemistry of State Oceanic Administration Third Institute of Ocea- nography State Oceanic Administration Xiamen 361005 China Third Institute of Oceanography State Oceanic Administration Xiamen 361005 China Chinese Arctic and Antarctic Administration Beijing 100860 China Department of Earth and Environmental sciences Rutgers University Newark NJ USA

The Prydz bay in the Antarctic is an important area in the Southern Ocean due to its unique geographic feature. It plays an important role in the carbon cycle in the Southern Ocean. To investigate the distributions of carbon dioxide in the atmosphere and surface seawater and its air-sea exchange rates in this region, the Chinese National Antarctic Research Expedition （CHINARE） had set up several sections in the Prydz bay. Here we present the results from the CHINARE-XVI cruises were presented onboard R/V Xue/ong from November 1999 to April 2000 and the main driving forces were discussed controlling the distributions of partial pressure of carbon dioxide. According to the partial pressure of carbon dioxide distributions, the Prydz bay can be divided into the inside and outside regions. The partial pressure of carbon dioxide was low in the inside region but higher in the outside region during the measurement period. This distribution had a good negative correlation with the concentrations of ehlorophyll-a in general, suggesting that the partial pressure of carbon dioxide was substantially affected by biological production. The results also indicate that the biological produetion is most likely the main driving force in the marginal ice zone in the Southern Ocean in summer. However, in the Antarctic divergence sector of the Prydz bay （about 64°S）, the hydrological processes become the controlling factor as the sea surface partial pressure of carbon dioxide is much higher than the atmospheric one due to the upwelling of the high DIC CDW, and this made the outside of Prydz bay a source of carbon dioxide. On the basis of the calculations, the CO2 flux in January （austral summer） was -3.23 mmol/（m^2 · d） in the inner part of Prydz bay, i.e. , a sink of atmospheric CO2, and was 0.62 mmol/（m^2 · d） in the outside part of the bay, a weak source of atmospheric CO2. The average air-sea flux of CO2 in the Prydz bay was 2.50 mmol/（m^2 · d）.

关键词： Prydz bay carbon flux carbon dioxide controlling factors

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Phylogenetic diversity of bacterial biofilms covering the settlement substrates of nona-porous abalones (Haliotis diversicolor supertexta)

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Acta Oceanologica Sinica 2009年第5期28卷 94-102页

作者： MA Ying WANG Zhiyong CHEN Zhengqiang LU Yu WANG Lin The Key Laboratory of science and Technology for Aquaculture and Food Safety Fisheries College Jimei University Xiamen 361021 China

The settlement substrates of nona-porous abalones （Haliotis diversicolor supertexta） are covered with biofilms in which several types of microorganisms coexist and interact. These microorganisms are usually important causes of juvenile abalone disease as well as organisms useful in promoting abalones’ adhesion. The bacterial community structure of the biofilms remains unclear. The aim of this research was to determine the genetic diversity and phylogenetic affiliation of the biofilm bacteria. Total DNA of bacteria in biofilms was extracted, and 16S rRNA gene clone library was constructed using the primers specific for the domain bacteria. Subsequently, 30 randomly selected positive clones were screened by PCR-restriction fragment length polymorphism （PCR-RFLP） analysis, and resulted in 15 different RFLP patterns. Sequences analysis of representatives from each unique RFLP type revealed high genetic diversity in the bacterial populations. These sequences fell into nine major lineages of the bacterial domains： α-, β-, γ-and δ-subdivisions of the Proteobacteria; Planctomycete, Actinobacteria, Firmicutes, V errucomicrobium spp., and CytophagaFlexibacter-bacteroides spp. Phylogenetic analysis indicated that the dominant phylotypes were most closely related to environmental and clinical burkholderia cepacia of the β-Proteobacteria, and Roseobacteria of the α-Proteobacteria.

关键词： nona-porous abalone (Haliotis diversicolor supertexta) settlement substrate bacterial biofilm 16S rRNA

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