一株枯草芽孢杆菌的分离鉴定、生物学特性及其对水质净化的作用

[背景]随着水产养殖业的发展和养殖集约化程度的提高,养殖水环境日趋恶化,养殖动物病害频发,而水产益生菌因其环境友好、安全而被广泛应用于水产养殖中.[目的]从南美白对虾养殖池底泥分离枯草芽孢杆菌,探究其体外生物学特性及对水质的净化作用,以期扩充微生态制剂的种质资源.[方法]采用稀释涂布平板法分离菌株,通过形态学观察、生理...     

黄菖蒲和美人蕉对水深梯度的响应差异

通过野外观测研究了黄菖蒲和美人蕉对水深梯度(10、30、50和70cm)的适应性。结果表明:随着水深的增加(1)黄菖蒲和美人蕉的分蘖数显著减少(P<0.05),但对株高、叶长和叶宽的影响不大;(2)2种植物叶片的叶绿素含量和根系活力均逐渐降低,叶片的丙二醛含量则逐渐升高,其中美人蕉的叶绿素含量、根系活力和叶片丙二醛含量的差异显著(P<0.05),而黄菖蒲仅根系活力的差异显著(P<0.05);(3)叶绿素荧光参数中,PSⅡ的最大光能转换效率(Fv/Fm)、表观电子传递速率(ETR)和光化学淬灭(qP)均呈显著下降的趋势(P<0.05),非光化学淬灭(qN)则显著增加(P<0.05);相对电子传递速率(rETR)及潜在最大电子传递速率(rETRm)、对光能的利用效率(α)和对强光的耐受能力(Ik)也都表现出明显的下降;美人蕉在10cm水深条件下各指标均高于黄菖蒲,但在水深较大的条件下总体上低于黄菖蒲。可见,水深较大时对黄菖蒲和美人蕉的生长产生了抑制作用,且对后者的抑制程度要大于前者。因此,在选择黄菖蒲和美人蕉进行湿地植物修复时,应保持浅水条件以利于其生长,其中黄菖蒲适应的水深(30～70cm)大于美人蕉(10cm...     

生物浮岛与漂浮植物对开放池塘水质净化效果

采用自主研发流量可控的密闭漂浮性水槽开展河道原位模拟实验, 研究了挺水植物菖蒲(种植于浮岛上)与漂浮植物凤眼莲对受到城市污水污染的开放池塘水质净化效果以及系统中氮磷等污染物的归趋。结果表明, 菖蒲和凤眼莲对水体藻类密度和叶绿素a削减率达到90%以上, 对COD_Mn浓度削减率达到45%以上。经过10 m长水槽后, 种植了凤眼莲的水槽水体其TN和TP浓度分别由3.71和0.24 mg/L降低至1.71和0.09 mg/L, 而设置有菖蒲浮岛的水槽其水体TN和TP浓度则分别降低至2.69和0.16 mg/L。在水体N、P的总削减量中, 凤眼莲吸收作用分别占84.31%和77.52%, 而在菖蒲浮岛系统中, 菖蒲的吸收作用仅分别占7.72%和8.55%, 菖蒲净化系统中氮、磷的物理沉淀量显著高于凤眼莲组, 分别达到35.26%和51.58%, 但仍有57%和39%以上的氮和磷去向未知, 推测可能与浮岛上生长的生物膜有关。研究结果可为选用凤眼莲和浮岛植物修复技术进行污染水体生态修复理论研究与实践运用提供借签和参考。     

黄菖蒲适生环境筛选

对黄菖蒲（Iris pseudacorus L．）在旱生（CK）、湿生及水深10、30、60、100和150cm 7个不同栽培环境下的株高、叶干重、通气组织及部分生理指标的变化进行了比较分析。发现从旱生至30cm水深环境中,黄菖蒲的株高和叶干重均逐渐增加,而在水深60cm至150cm的环境中则呈降低趋势。在水深30cm的环境中处理40d,黄菖蒲叶片通气道与横截面积比及叶片SOD活性显著高于对照和其他处理组;相对电导率及脯氨酸含量随水深及处理时间的增加基本呈上升趋势。结果表明,黄菖蒲在水深约30cm的环境中生长状况最佳;在旱生、湿生和水深10cm的环境中能够生长良好;在60cm水深环境生长比较适宜;水深超过60cm,黄菖蒲不能正常生长。     

菰和菖蒲对富营养化水体净化效率的比较

研究了菰〔Zizania latifolia(Griseb.)Stapf〕、菖蒲(Acorus calamusL.)及它们的复合体系对富营养化水体的净化效果。结果表明,菰和菖蒲在供试富营养化水体中均能正常生长,二者单独种植体系或等量混合种植体系对富营养化水体均有一定的净化能力。单独种植的菰和菖蒲及二者的混合种植体系对供试水体中总氮的去除率分别为92.8%、92.7%和94.9%;对氨氮的去除率分别为95.5%、97.4%和96.6%;对总磷的去除率分别为83.9%、94.3%和84.7%;对CODCr的去除率分别为83.0%、85.5%和86.7%。单独种植的菖蒲对总磷的去除效果明显好于单独种植的菰和二者的混合种植体系。菰和菖蒲的混合种植体系对富营养化水体的净化效果与单独种植体系无明显差异。     

Cd-Cu复合胁迫对黄菖蒲叶片及根系中Cd和Cu的积累及其迁移率的影响

采用水培法研究了Cd-Cu复合胁迫(5 mg·L-1Cd-10 mg·L-1Cu;5 mg-L-1Cd-20 mg·L-1Cu;25 mg·L-1 Cd-10 mg·L-1Cu;25 mg·L-1Cd-20 mg·L-1Cu)条件下黄菖蒲(Iris pseudacorus L.)叶片和根系对Cd和Cu的积累作用及黄菖蒲体内Cd和Cu迁移率的变化规律.结果表明,在50 d的胁迫期内,黄菖蒲叶片和根系中的Cd和Cu含量分别随培养液中Cd和Cu浓度的提高而增加,并随胁迫处理时间的延长基本呈增加的趋势,在胁迫末期明显提高.在不同浓度Cd-Cu复合胁迫条件下,黄菖蒲根系中的Cd和Cu积累量明显高于叶片.在Cd浓度不变的条件下,黄菖蒲叶片和根系中的Cd含量随培养液中Cu浓度的提高而增加;在Cu浓度不变的条件下,黄菖蒲叶片中Cu含量随培养液中Cd浓度的提高而降低.在不同胁迫时间,黄菖蒲植株对Cd的迁移率不同,但培养液中Cd的浓度较高,黄菖蒲植株对Cd的迁移率也较高;黄菖蒲植株对Cu的迁移率随胁迫时间的延长基本上呈上升趋势.研究结果显示,在Cd-Cu复合胁迫条件下,Cu对黄菖蒲体内Cd的吸收具有一定的协同吸收作用,而Cd对Cu的吸收则有一定的拈抗作用.     

Cd胁迫对黄菖蒲幼苗4种抗氧化酶活性的影响

采用水培法对Cd胁迫下黄菖蒲（Iris pseudacorus L．）幼苗叶片和根系中超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）及抗坏血酸过氧化物酶（APX）的活性进行了研究。结果表明,10、40和120mg·L^-1Cd胁迫下,黄菖蒲幼苗叶片和根系中4种酶活性的变化不同。10和40mg·L^-1 cd胁迫下,黄菖蒲幼苗叶片和根系中的POD及APX活性、叶片中的SOD活性及根系中的CAT均明显高于对照;在120mg·L^-1 Cd胁迫下,叶片中的POD活性及根系中的POD和CAT活性均高于对照;各处理组根系中的SOD活性均低于对照。随处理时间的延长,40和120mg·L^-1Cd胁迫处理组叶片的CAT活性和120mg·L^-1Cd胁迫处理组根系的APX活性逐渐降低,其他处理组不同酶的活性逐渐升高或先升后降。黄菖蒲叶片及根系中的4种酶对Cd胁迫的响应能力有差异,其中POD可能是黄菖蒲耐Cd胁迫的主要抗性诱导酶。     

菖蒲和石菖蒲的生长特性及其对生活污水净化功能的比较研究

以菖蒲和石菖蒲为对象,通过漂浮栽植研究了其在不同浓度生活污水中的生长特性及其对污水TN、NH3-N、TP、CODCr的去除情况。结果表明,菖蒲在中浓度污水中生长最好,其生物量显著高于低、高浓度,而石菖蒲在低浓度污水中生长最好,其生物量显著高于中、高浓度。菖蒲对中浓度污水中TN、TP、CODCr的去除率分别为96.85%、82.62%、88.89%,对低浓度污水中TN、TP、CODCr的去除率分别为99.87%、91.38%、83.34%,净化效果均高于高浓度(96.58%、80.96%、77.55%),但对NH3-N的去除率在高浓度污水中最好,为99.46%。石菖蒲净化中浓度污水的效果最好,对TN、NH3-N、TP、CODCr的去除率分别为94.20%、78.06%、89.43%、74.98%,对高浓度污水中的NH3-N、TP、CODCr的去除效果也较好,去除率分别为94.62%、91.95%、87.08%,但对TN去除率为59.14%,显著低于低浓度(98.73%)和中浓度(94.20%)。     

水生植物对富营养水体水质净化作用研究

利用富营养浅水湖泊武汉东湖中所建立的大型实验围隔系统,对沉水植物的水质净化作用作了现场实验研究。重建后的沉水植物可以显著改善水质,水体透明度显著提高,水色降低。在研究期间,水生植物围隔CODcr和BOD5一般分别为20和5mg/L左右;对照围隔和大湖水体则分别约为40和10mg/L。水生植物围隔水体中检出的有机污染种类也较对照围和大湖水体柢。实验结果表明恢复以沉水植物为主的水生植被是改善营养湖泊水质和重建生态系统的有效措施。     

水生植物对水体水质的净化作用分析

随着中国工业化进程的推进,近年来,水生植物对水体水质的净化作用的研究需求越来越急切。水生植物对水体水质起主要净化作用,本文主要通过对水生植物的概念,水生植物对水体水质净化作用的应用,以及水生植物对水体水质的净化原理三个方面的阐述,从而对水生植物对水体水质的净化作用进行分析。     

马蔺IlWRKY28基因的克隆与表达分析

马蔺(Iris lactea var. chinensis)是鸢尾属多年生草本盐生植物,具有很高的耐盐性和观赏价值。为研究马蔺耐盐的分子机制,通过cDNA末端快速扩增技术(RACE)从马蔺中克隆到一个WRKY转录因子基因IlWRKY28,获得了1 302 bp的全长cDNA序列,其包含一个108 bp 5′末端非翻译区(UTR),一个174 bp 3′末端UTR和一个1 020 bp开放阅读框。IlWRKY28编码339个氨基酸,预测的蛋白质分子量为37.22 kD,等电点为7.04。氨基酸序列分析显示,IlWRKY28包含一个保守的WRKY基序和一个C2H2型锌指结构域。系统发育分析表明,马蔺IlWRKY28与菠萝(Ananas comosus)AcWRKY28和藏北嵩草(Kobresia littledalei)ClWRKY28亲缘关系最近。荧光定量PCR分析显示,盐处理后,IlWRKY28基因在马蔺地上部显著上调表达。该研究结果为进一步研究IlWRKY28在马蔺适应高盐胁迫中的功能和作用机制奠定了重要的分子基础。     

大叶桉叶水浸提液成分分析

为进一步明确大叶桉的化学成分,对大叶桉叶水浸提液分别用不同极性的有机溶剂石油醚、乙酸乙酯和正丁醇进行萃取,对各萃取相进行GC-MS分析。结果表明:大叶桉叶水浸提液共含有37种化合物,其中,石油醚萃取相中含有20种,主成分为草酸丁基异己酯(37.24%);乙酸乙酯萃取相中含有16种,主成分为2,2-二亚甲基双[6-(1,1-二甲基乙基-4-甲基)]-苯酚(50.05%);正丁醇萃取相中含有5种,主成分为丙基-2-甲基丁酸酯(54.57%)。在所有成分中,酯类物质居多,也有少量的烯、酮、醇、苯和烷烃。1-甲基,4-(1-甲基乙基)-1,4环己二烯、2,2-二亚甲基[6-(1,1-二甲基乙基)-4-甲基]苯酚、1-十八烯和二十烷为石油醚和乙酸乙酯的共有成分;1、2-苯二甲酸单(2-乙基己基)酯为乙酸乙酯和正丁醇的共有成分。该研究进一步明确了大叶桉的化学成分,为其在医药、化工和化感方面的应用研究奠定了基础。     

Rearing of larvae of Anopheles stephensi, using water replacement, purification and automated feeding

For rearing progenies of single females of Anopheles stephensi (Liston) (Diptera: Culicidae), a method is proposed using small trays with perforated side walls (called open trays) to allow for water replacement and purification using an aquatic plant Vallisneria spiralis. The performance of this method has been compared with that of rearing single progenies in trays without mesh side walls (closed trays). Effects of larval density and food quantity on rearing performance have been studied in both types of trays. In open trays the larvae developed about 2 days faster than in closed trays. Male larvae developed significantly faster (0.4 days) than females, but this difference was not significantly influenced by method of rearing or larval density. Adults from fast developing larvae were heavier than those emerging from slowly developing larvae from a single egg batch. Open trays gave higher larval survival than closed trays. A system has been developed to automatize larval feeding.

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Elevage des larves issues de femelles isolées d' Anopheles stephensi Liston (Dipt.; Culicidae), avec utilisation d'un système automatique d'alimentation et de renouvellement de l'eau

Résumé Une technique, utilisant des récipients avec des parois latérales perforées (baptisés récipients ouverts) permettant le renouvellement de l'eau et sa purification par une plante aquatique (Vallisneria piralis), est préconisée pour élever la descendance de femelles isolées de A. stephensi. Les performances de cette technique ont été comparées à l'élevage dans des récipients sans parois perforées (récipients clos). L'influence de la densité et de la quantité d'aliments a été examinée dans les deux cas. Dans les récipients ouverts les larves se développent plus vite (deux jours de moins); les mâles se développent plus vite que les femelles, mais cette différence (0,4 j) n'est significiativement pas influencée par les techniques d'élevage ou la densité. Pour une même ponte les adultes provenant des larves se développant le plus vite sont plus gros que ceux qui sont issus des larves les plus lentes. Le récipient ouvert donne une meilleure survie. Un système a été mis au point pour automatiser l'alimentation.

     

Testing the adaptive plasticity of Iris pumila leaf traits to natural light conditions using phenotypic selection analysis

A multivariate selection analysis has been used to test the adaptiveness of several Iris pumila leaf traits that display plasticity to natural light conditions. Siblings of a synthetic population comprising 31 families of two populations from contrasting light habitats were grown at an open dune site and in the understory of a Pinus nigra stand in order to score variation in phenotypic expression of six leaf traits: number of senescent leaves, number of live leaves, leaf length, leaf width, leaf angle, and specific leaf area. The ambient light conditions affected the values of all traits studied except for specific leaf area. In accordance to ecophysiological expectations for an adaptive response to light, both leaf length and width were significantly greater while the angle between sequential leaves was significantly smaller in the woodland understory than at the exposed dune site. The relationship between leaf traits and vegetative fitness (total leaf area) differed across light habitats as predicted by functional hypotheses. The standardized linear selection gradient (β′) for leaf length and width were positive in sign in both environments, but their magnitude for leaf length was higher in the shade than under full sunlight. Since plasticity of leaf length in the woodland shade has been recognized as adaptive, fitness cost of producing plastic change in leaf length was assessed. In both of the available methods used, the two-step and the multivariate regression procedures, a rather high negative association between the fitness value and the plasticity of leaf length was obtained, indicating a cost of plasticity. The selection gradient for leaf angle was weak and significant only in the woodland understory. Genetic correlations between trait expressions in contrasting light environments were negative in sign and low in magnitude, implying a significant genetic variation for plasticity in these leaf traits. Furthermore, leaf length and leaf width were found to be genetically positively coupled, which indicates that there is a potential for these two traits to evolve toward their optimal phenotypic values even faster than would be expected if they were genetically independent.     

The effect of two biological control agents, the weevil Neochetina eichhorniae and the mirid Eccritotarsus catarinensis on water hyacinth, Eichhornia crassipes , grown in culture with water lettuce, Pistia stratiotes

We assessed the effect of two biological control agents, the mirid Eccritotarsus catarinensis (Carvalho) and the weevil Neochetina eichhorniae (Warner), singly or in combination, on the competitive ability of their host plant, water hyacinth, Eichhornia crassipes (Mart.) Solms-Laub., grown in a screen house, in competition with another aquatic plant (Pistia stratiotes L.). Water hyacinth plant growth characteristics measured included fresh weight, leaf and petiole lengths, number of inflorescences produced, and new shoots. Without herbivory, water hyacinth was 18 times more competitive than water lettuce (across all experimental combinations of initial plant densities), as estimated from fresh weights. Both insect species, singly or in combination, reduced water hyacinth plant growth characteristics. E. catarinensis alone was less damaging than the weevil and under normal conditions, i.e., floating water hyacinth, is not expected to increase control of water hyacinth beyond that of the weevil. When combined with the weevil, half the inoculum of weevils and half the inoculum of mirids produced the same growth reduction as the full inoculum of the weevil. Under conditions where the weevils are not effective because water hyacinths are seasonally rooted in mud, the mirid, which lives entirely on leaves, should become a useful additional biological control agent. Handling Editor: John Scott.     

The effect of Euglena viridis on immune response of rohu, Labeo rohita (Ham.)

The study evaluated the effect of dietary doses of Euglena viridis on the immune response and disease resistance of Labeo rohita fingerlings against infection with the bacterial pathogen Aeromonas hydrophila. L. rohita fingerlings were fed with diet containing 0 (Control), 0.1 g, 0.5 g, 1.0 g Euglena powder kg⁻¹ dry diet for 90 days. Biochemical (serum total protein, albumin, globulin, albumin:globulin ratio), haematological (WBC, RBC, haemoglobin content) and immunological (superoxide anion production, lysozyme, serum bactericidal activity) parameters of fish were examined after 30, 60 and 90 days of feeding. Fish were challenged with A. hydrophila 90 days post-feeding and mortalities were recorded over 10 days post-infection. The results demonstrate that fish fed with Euglena showed increased levels of superoxide anion production, lysozyme, serum bactericidal activity, serum protein and albumin (P < 0.05) compared with the control group. Following challenge with A. hydrophila less survivability was observed in the control group (56.65%) than the group fed the experimental diets. The group fed 0.5 g Euglena kg⁻¹ dry diet showed the highest percentage survival (75%). These results indicate that Euglena stimulates the immunity and makes L. rohita more resistant to A. hydrophila infection.     

Factors affecting entomopathogenic nematode infection ofPlutella xylostella on a leaf surface

We investigated the ability of entomopathogenic nematodes to infect diamondback moth (DBM),Plutella xylostella (L.) (Lepidoptera: Plutellidae) on a leaf surface. In a leaf disk assay, mortality of late stage DBM larvae ranged from <7% caused bySteinernema kushidai Mamiya to >95% caused byS. carpocapsae (Weiser) All strain. LC₅₀ values forS. carpocapsae, S. riobravis Cabanillas, Poinar & Raulston, andHeterorhabditis bacteriophora Poinar NC1 strain were 14.6, 15.4, and 65.4 nematodes/larva, respectively.S. carpocapsae, S. riobravis, andH. bacteriophora caused 29%, 33%, and 14% mortality of DBM pupae, respectively. DBM mortality caused byS. carpocapsae on radish declined at low (<76%) to moderate (76–90%) RH, because nematode survival and infectivity declined at low (<76%) to moderate (76–90%) RH. However, DBM mortality caused byS. riobravis did not decline with RH.S. riobravis survival declined with RH, but infectivity did not. Overall, nematode survival and infectivity to DBM larvae were lower forS. riobravis than forS. carpocapsae. In addition, DBM mortality was higher on radish plants (pubescent leaves) than on cabbage plants (glaborous leaves).     

Pilot-scale production and purification of a staphylokinase-based fusion protein over-expressed in Escherichia coli

SFH, a recombinant staphylokinase-based fusion protein linked by the factor Xa recognition peptide at the N-terminus of hirudin, is a promising therapeutic candidate for thromboembolic diseases. To develop SFH into a new thrombolytic agent, scaled-up production was carried out to provide sufficient preparation for animal safety and clinical studies. Here, we describe a pilot-scale cultivation and purification process for the production of SFH. A high-cell-density fed-batch cultivation for the production of SFH in E. coli was developed in a 40-L bioreactor, which produced about 1.1 g/L of recombinant protein. SFH was purified to homogeneity from the E. coli lysate by expanded bed adsorption chromatography and anion-exchange chromatography, with over 99% purity and 54% recovery. Moreover, the residual endotoxin content was less than 0.5 EU/mL. The molecular weight and in vitro bioactivity of SFH were also determined by electrospray ionization-mass spectrometry (ESI-MS) and fibrinolytic activity assay, respectively.     

Biological activity of maize leaf extracts against the African armyworm,Spodoptera exempta

Maize (Zea mays L.) leaf tissue of cv Bastille and cv Michoacan 12 was extracted with n-hexane. The extracts were bioassayed against 5th instar African armyworm,Spodoptera exempta (Walker)(Lepidoptera: Noctuidae), by feeding the larvae on agar based media or sucrose impregnated glass fibre discs. The hexane extract of the ‘resistant’ cv Bastille exhibited feeding deterrency and toxicity which were not shown by the ‘susceptible’ cv Michoacan 12. The hexane extract of cv Bastille was adsorbed onto silica gel, the solution filtered off and the adsorbed component taken up into ethyl acetate. Bioassay of these fractions indicated that the toxic and deterrent action was retained in the ethyl acetate fraction. Preparative thin layer chromatography of the ethyl acetate fraction isolated two biologically active constituents. These were both growth inhibitors and lethal by ingestion to the 5th instar African armyworm. Implications for resistance in maize varieties to insect pests are discussed.