Study on the Quantity Dynamic Changes of Heterobacteria and Vibrios in Larvae Industrialized Culture System

[Objective] The dynamic change of heterobacteria and vibrios in larvae industrialized culture system was studied to provide scientific reference for healthy cultivation of shrimp.[Method] The heterobacteria,vibrios and pathogenic vibrio parahaemolyticus were monitored in larvae industrialized culture system.[Result] The heterobacteria,vibrios and pathogenic vibrio parahaemolyticus were the most in fertilized eggs of shrimp but the least in nauplius,then their number would increase with growth. During whole rearing period,both heterobacteria in larvae,vibrios in water would increase by one order of magnitude,while both vibrios in larvae and heterobacteria in water would increase by two orders of magnitude.There were many heterobacteria and vibrios but few vibrio parahaemolyticus in living bait.The correlation coefficients between larvae and heterobacteria and vibrios in water were 0.704 and 0.840 in culture system respectively,while the correlation among heterobacteria,vibrios in living bait and larvae,water were weak or negative.[Conclusion] There was a dynamic relation between water and larvae in rearing period,and restrictly control of culture condition would restrain the occurrence of disease caused by vibrio parahaemolyticus,besides that bacteria number in bait was not obviously correlated with bacteria nubmer in culture system.     

苍耳提取液与植物内生真菌抗菌活性研究(英文)

[Objective] The study was aimed to provide theoretical foundation for developing a new fungicide with high activity,low toxicity and low residua.[Method] Based on the plant growth rate and inhibition zone methods,the antifungal activity of extracts of every organ of Xanthium sibiricum,the fermentation liquid of endophytic fungi MC4-23,and the extract of mycelium were tested with six kinds of pathogenic fungi and three kinds of pathogenic bacteria respectively.[Result] The extract of every organ of Xanthium sibiricum,the fermentation liquid of endophyic fungi MC4-23,and the extract of mycelium all had antifungal activities.The acetone extracts of Xanthium sibiricum had stronger antifungal activities than the ethanol extracts of Xanthium sibiricum.The extracts of leaf have the strongest antifungal activities against fungi and bacteria.The inhibition rates of the six pathogenic fungi were all above 47.8%,and the inhibition diameters of the three pathogenic bacteria were all above 0.5 centimeter.The fermentation liquid of endophytic fungi also had strong activities against the fungi and bacteria.The inhibition rates of the six pathogenic fungi were all above 68.1%.The extracts of root of Xanthium sibiricum and the mycelium of endophytic fungi were all had antifungal activities.[Conclusion] Xanthium sibiricum and endophyic fungi MC4-23 had great value of development and utilization.     

苍耳提取液与植物内生真菌抗菌活性研究(英文)

[Objective] The study was aimed to provide theoretical foundation for developing a new fungicide with high activity,low toxicity and low residua.[Method] Based on the plant growth rate and inhibition zone methods,the antifungal activity of extracts of every organ of Xanthium sibiricum,the fermentation liquid of endophytic fungi MC4-23,and the extract of mycelium were tested with six kinds of pathogenic fungi and three kinds of pathogenic bacteria respectively.[Result] The extract of every organ of Xanthium sibiricum,the fermentation liquid of endophyic fungi MC4-23,and the extract of mycelium all had antifungal activities.The acetone extracts of Xanthium sibiricum had stronger antifungal activities than the ethanol extracts of Xanthium sibiricum.The extracts of leaf have the strongest antifungal activities against fungi and bacteria.The inhibition rates of the six pathogenic fungi were all above 47.8%,and the inhibition diameters of the three pathogenic bacteria were all above 0.5 centimeter.The fermentation liquid of endophytic fungi also had strong activities against the fungi and bacteria.The inhibition rates of the six pathogenic fungi were all above 68.1%.The extracts of root of Xanthium sibiricum and the mycelium of endophytic fungi were all had antifungal activities.[Conclusion] Xanthium sibiricum and endophyic fungi MC4-23 had great value of development and utilization.     

Preliminary Studies on Antifungal Activity of Xanthium sibiricum and the Endophytic Fungi

[Objective] The study was aimed to provide theoretical foundation for developing a new fungicide with high activity,low toxicity and low residua.[Method] Based on the plant growth rate and inhibition zone methods,the antifungal activity of extracts of every organ of Xanthium sibiricum,the fermentation liquid of endophytic fungi MC4-23,and the extract of mycelium were tested with six kinds of pathogenic fungi and three kinds of pathogenic bacteria respectively.[Result] The extract of every organ of Xanthium sibiricum,the fermentation liquid of endophyic fungi MC4-23,and the extract of mycelium all had antifungal activities.The acetone extracts of Xanthium sibiricum had stronger antifungal activities than the ethanol extracts of Xanthium sibiricum.The extracts of leaf have the strongest antifungal activities against fungi and bacteria.The inhibition rates of the six pathogenic fungi were all above 47.8%,and the inhibition diameters of the three pathogenic bacteria were all above 0.5 centimeter.The fermentation liquid of endophytic fungi also had strong activities against the fungi and bacteria.The inhibition rates of the six pathogenic fungi were all above 68.1%.The extracts of root of Xanthium sibiricum and the mycelium of endophytic fungi were all had antifungal activities.[Conclusion] Xanthium sibiricum and endophyic fungi MC4-23 had great value of development and utilization.     

Initial Study on the Antibacterial Action of Mentha spicata Linn Extract in Vitro

[Objective] The research aimed to study the antibacterial action of Mentha spicata Linn extract in vitro.[Method] The cylinder-plate method was used to measure the inhibitory zone size of Mentha spicata Linn extract.The test-tube double dilution method was used to measure the minimal inhibitory concentration（MIC）of Mentha spicata Linn extract on the four kinds of animal pathogenic bacteria,which could determine the antibacterial effect.[Result] The decoction,alcohol extract and volatile oil of Mentha spicata Linn had the different antibacterial effects on the four kinds of animal pathogenic bacteria.Especially the inhibition effect on the pig staphylococcus was comparatively obvious.MIC of Mentha spicata Linn decoction I,alcohol extract III,volatile oil V and volatile oil VI were respectively 31.25,62.5,2.32 and 2.31 mg/ml.[Conclusion] The decoction,alcohol extract and volatile oil of Mentha spicata Linn had the certain antibacterial effect in vitro,and the extracts which were gained by the different extraction methods had the different inhibition effects on the experimental bacteria.     

“胰岛素-转铁蛋白-硒钠”对山羊卵母细胞体外成熟及胚胎发育的影响(英文)

[Objective] The research aimed to enhance culture efficiencies of oocyte and embryo of goat in vitro and to explore serum-free culture system in vitro.[Method] At present,the conventional solutions of oocyte maturation and embryo development in vitro were always added into 1% ITS(Insulin-transferrin-selenium) or using 1% ITS to replace FBS in 2 kinds culture solutions for conducting in vitro cultures of goat oocyte and parthenogenetic embryo.The influences of ITS on their developments were detected.[Result] ITS in maturation liquid of oocytes could not increase oocytes maturation rate but significantly increased blastocyst rate (58.06% vs. 48.19%)of parthenogenetic embryo.If FBS in maturation liquid of oocytes was replaced by ITS, the maturation rate, cleavage rate and blastocyst rate were basically unchanged.Adding ITS into embryo medium could increase blastocyst rate (68.30% vs. 56.82%)of parthenogenetic embryo of goat.If FBS in embryo medium was replaced by ITS,the cleavage rate didn’t change basically,while the blastocyst rate in ITS was obviously lower than that in FBS group(42.33% vs.56.82%).[Conclusion] ITS could promote maturation of oocyte in vitro and early embryonic development, in addition,ITS could replace serum in maturation medium of oocyte as serum-free culture system for conducting relevant researches.     

Effect Evaluation about Pathogenic Bacteria of Diary Cow Mastitis Treated with Ruyuankang

In this research,36 cows with clinical mastitis were treated with the Chinese herb injection Ruyuankang which was developed by our group.Milk samples of pre and post treatment of Ruyuankang were collected for isolation and characterization of pathogenic bacteria.The dynamic changes of pathogenic bacteria were investigated as well.The results indicated that Ruyuankang could effectively inhibit the common pathogenic bacteria of mastitis which result in the negative rate of pathogenic bacteria 71.74%.     

丁草胺微乳液的研究(英文)

[Objective] The properties of butachlor microemulsion were studied for developing a new formulation of new pesticides.[Method]AT method was used to determine the pseudo-ternary phase diagram to confirm the formulation of butachlor microemulsion.Through the measurement of electrical conductivity,the W/O and O/W types in microemulsion region of butachlor/emulsifier/water system were confirmed and the change of phase behavior during preparation process was discussed.[Result]The dilution stability,low temperature stability,heat stability,ageing stability,density and viscosity etc.of butachlor microemulsion met the requirement of the experiments,which demonstrated the qualified quality of butachlor.The density decreased linearly with the increase of temperature and the change of viscosity with temperature conformed to Andrade equation.[Conclusion]The research was helpful to the application of butachlor microemulsion in pesticide formulation.     

Character of viral clones in serial passage of a nuclear polyhedrosis virus in vitro

In this paper,the character of viral clones from early and late passages after serial passages of Trichoplusia ni single nuclear polyhedrosis virus in a Tn 5B1-4 cell line is described.It demonstrated that no significant difference was observed in the infectivity of the cell culture supernatants of various passages to the cell line.The number of polyhedra produced in a cell and infectivity of polyhedra to T.ni larvae declined strikingly with the increase of passages.The polyhedra without virions began to increase from passage to passage.The result of restriction enzyme digestion showed that the DNA restriction fragments of the clones were different from wild virus DNA,although they came from a homogeneous viral DNA.The mutation of viral DNA resulted in the in crease of noninfectious polyhedra without virions and in the increase of the number of polyhedra produced in cell line as well as virulence of the polyhydrosis inclusion bodys to T.ni larvae after prolonged passages of Tn SNPV in the cell culture.     

百蕊草无性系建立与瓶外生根研究(英文)

[Objective] The aim of this study is to establish the rapid micro-propagation system in Thesium chinense Turcz.[Method]With stem fragments of wild Thesium chinense Turcz as explants,different culture media were designed to conduct induction culture,strengthening plantlet culture and in vitro rooting.[Result]The optimum medium for inducing clustered shoots was determined to be MS medium appended with 1.5 mg/L 6-BA,0.01 mg/L NAA and 0.3 mg/L 2,4-D;in addition,60 mg/kg ABT was suitable for rooting,by which the percentage of rooted plantlets reached 76.6%.[Conclusion]This study simplified the procedures of tissue culture in Thesium chinense Turcz and enhanced the proliferation rate,providing basis for artificial cultivation and resource protection of Thesium chinense Turcz.     

工厂化育苗养殖体系内细菌数量动态变化研究

以异养菌、弧菌、副溶血弧菌为监测对象,对中国对虾的工厂化育苗进行实时监测,探寻育苗过程中细菌的动态变化规律,为对虾健康养殖提供科学依据。     

Study on the Quantity Dynamic Changes of Heterobacteria and Vibrios in Larvae Industrialized Culture System

[Objective] The dynamic change of heterebacteria and vibrios in larvae industrialized culture system was studied to provide scientific reference for healthy cultivation of shrimp. [Method] The hcterobacteria, vibrios and pathogenic vibrio parahaemolyticus were monitored in larvae industrialized cul-ture system. [Result] The heterobacteria, vibrios and pathogenic vibrio parahaemolyticus were the most in fertilized eggs of shrimp but the least in naupli-us, then their number would increase with growth. During whole rearing period, both heterobscteria in larvae, vibrios in water would increase by one order of magnitude, while both vibrios in larvae and heterobacteria in water would increase by two orders of magnitude. There were many heterohacteria and vibri-os but few vibrio parahaemolyticus in living bait. The correlation coeffcients between larvae and heterobacteria and vibrios in water were 0.704 and 0.840 in culture system respectively, while the correlation among heterobacteria, vibrios in living bait and larvae, water were weak or negative. [Conclusion] There was a dynamic relation between water and larvae in rearing period, and restrictly control of culture condition would restrain the occurrence of disceae caused by vibrio parahaemolyticus, besides that bacteria number in bait was not obviously correlated with bacteria nubmer in culture system.     

工厂化育苗养殖体系内细菌数量动态变化研究

[目的]探寻工厂化育苗中细菌的动态变化规律,为对虾健康养殖提供科学依据。[方法]对中国对虾育苗期养殖体系中的异养菌、弧菌、致病性副溶血弧菌进行监测。[结果]异养菌、弧菌和致病性副溶血菌数量都是对虾受精卵中高,无节幼体中最低,而后逐渐升高。在整个育苗期,对虾幼体中异养菌和养殖水体中弧菌增加1个数量级,对虾幼体中弧菌和养殖水体中异养菌均增加2个数量级。活饵中异养菌和弧菌数量很大,致病性副溶血弧菌量很低。养殖体系中幼体与水体中的异养菌和弧菌的相关系数分别为0.704和0.840;活饵中异养菌、弧菌与对虾幼体和养殖水体相关性很低或呈负相关。[结论]育苗期养殖水体与对虾幼体中细菌数量变化具有动态联系,严格控制养殖条件副溶血弧菌很难引起幼体疾病爆发,饵料中细菌数量与养殖系统中细菌数量无明显相关性。     

文蛤生物体及内脏中弧菌的研究

[目的]鉴定文蛤体内弧菌的类型及致病性强度,并筛选治疗这些病原菌的最佳药物。[方法]用TCBS培养基从文蛤生物体内分离出12株弧菌,对其进行致病性试验,对致病性较强的3株弧菌进行鉴定及药物敏感试验。[结果]该3株菌分别为河流弧菌H04（Vibrio fluvialis）,H06副溶血弧菌（Vibrio parahaemolyticus）,H11创伤弧菌（Vibrio vulnificus）,其对环丙沙星、先锋赛高度敏感,对青霉素G、先锋霉素V有耐药性,临床上可以首选环丙沙星、先锋赛作为治疗这些病原菌的药物,其次是头孢曲松、氟哌酸、丁胺卡那、四环素、庆大霉素等。[结论]结果为文蛤疾病的防治提供了理论依据。     

副溶血性弧菌外膜蛋白的提取与免疫鉴定

[目的]为进一步确定致病性副溶弧菌共有的特异性抗原和保护性抗原奠定基础。[方法]通过小鼠毒力试验研究5株副溶血性弧菌菌株对小鼠的致病性,比较在不同培养基和培养时间下所提取的外膜蛋白的SDS-PAGE图谱并通过Western blotting分析研究副溶血性弧菌菌株的免疫原性。[结果]5株临床分离的副溶血性弧菌菌株明显对小鼠具有不同的致病性,同时能在兔血平板上形成明显的溶血圈,为TDH阳性菌株。通过菌体免疫获得的多克隆抗体,与除副溶血性弧菌外的其他试验菌株均无交叉反应。Western blotting分析显示该多克隆抗体与5株致病性副溶血弧菌菌株可发生程度不等的阳性反应,提取的外膜蛋白具有较好的免疫原性。[结论]该研究为制备有效预防副溶血性弧菌引起的疾病的菌苗提供了理论基础。     

致病性副溶血性弧菌特异性三重PCR检测方法研究

[目的]建立同时检测致病性副溶血性弧菌gyrase、tdh、toxR基因的三重PCR快速检测方法。[方法]用3种基因的特异性引物分别对副溶血性弧菌ATCC33847的模板DNA进行单一扩增,找到各自引物最佳扩增条件;再用3种引物同步对模板DNA进行扩增,通过优化引物浓度、引物间比例以及退火温度,建立最佳扩增体系。[结果]在最佳三重PCR反应条件下,gyrase、tdh和toxR能同时扩增出清晰条带,大小分别为91、269和368 bp。[结论]该研究为致病性副溶血性弧菌的快速检测提供了一种新的技术方法。     

利用PCR技术快速检测水产品中副溶血性弧菌

[目的]对副溶血性弧菌的检测进行研究。[方法]采用聚合酶链式反应（PCR）,以标准菌株（单增李斯特菌、哈氏弧菌）作为阴性对照对3株副溶血性弧菌进行特异性扩增,扩增引物采用副溶血性弧菌如基因中的tlh-3和tlh-4,同时利用PCR对人工污染的白对虾中的副溶血性弧菌进行检出限测定。[结果]结果表明,通过PER扩增,副溶血性弧菌在约449bp处扩增出特异性条带,单增李斯特菌、哈氏弧菌均未出现任何条带,扩增片段表现出极好的特异性。菌液稀释到3．3×10^2cfu／ml时,将其污染到白对虾中作PCR仍可扩增出目的片断。[结论]该研究表明PER检测方法快捷、特异性好、敏感性高,可以作为副溶血性弧菌辅助检测方法。     

硝化细菌在澳洲银鲈工厂化养殖中的应用初探

[目的]为硝化细菌在工厂化养殖中的应用提供科学依据。[方法]在工厂化养殖池中,研究了施放硝化细菌前后水体中的氨态氮、亚硝酸氮、溶氧量、化学耗氧量等水化指标的变化情况。[结果]在施放硝化细菌前,实验池氨态氮含量上升趋势与对照池大致相同,在施放硝化细菌后第4天氨态氮含量出现下降,2~3 d后又开始缓慢上升。亚硝酸氮变化趋势与氨态氮大致相同,实验池在施放硝化细菌后第9天亚硝酸氮含量开始缓慢回升。对照池和实验池的溶氧均呈下降趋势。对照池和实验池的化学耗氧量在施放硝化细菌前上升明显,施放硝化细菌后上升放缓。[结论]在工厂化养殖池中施放硝化细菌,能有效改善养殖环境。     

高位虾池水体细菌和弧菌的数量变化及影响因素

研究了位于广东湛江东海岛北寮村和庵里村的4个凡纳滨对虾（Penaeus vannamei）高位池养殖过程水体中的可培养异养细菌和弧菌的数量变化及其影响因素.结果表明：1）北寮村虾池BL1和BL2的可培养异养细菌数量变化均呈现中间高两头低、后期多于初期的单峰变化趋势;BL1的弧菌数量变化趋势和可培养异养细菌的变化趋势相似,而BL2则是前期稍微升高而后下降,后期又呈升高态势,峰值出现在养殖末期;水源地BL3的可培养异养细菌数量变化为初期平缓,后期明显多于初期,大体呈上升态势;而BL3弧菌数量变化趋势则比较平缓.2）庵里村发病的虾池AL1的可培养异养细菌和弧菌数量呈现出单峰变化趋势,在养殖中途爆发病害时,可培养异养细菌数量高出未发病的正常池2个数量级;庵里村虾池AL2的可培养异养细菌数量变化趋势也是中间高两头低,而弧菌的数量变化则是无规律的波动;水源地AL3的可培养异养细菌数量变化大体呈下降趋势,弧菌数量放养初期最高.3）高位虾池的可培养异养细菌和弧菌数量虽多于水源地,但差异不显著,同一个位点,细菌和弧菌数量呈现同步增多或减少.虾场水体的可培养异养细菌和弧菌数量与多数环境因子之间相关性不显著,这与虾池养殖生态系统受人为因素影响较大有关,同时反映了高位虾池生态系统受多种因子作用的复杂性.