Modification of Amino-acid Concentrations Induced by Mycoplasmas in Cell Culture Medium

BOTH fermenting and non-fermenting mycoplasmas induce chromosomal aberrations in cultured cells^1–4, often accompanied with changes in cell morphology⁴. Aula and Nichols suggested that such aberrations are induced by depletion of the essential amino-acid arginine from culture media³, since non-fermenting mycoplasmas utilize it as an energy source⁵. Furthermore, media prepared without arginine or other essential amino-acids cause extensive chromosome damage to certain cultured cells⁶. We have investigated the possibility that such changes in the cell-free media could produce such anomalies in cultured cells.     

Long-Term Survival of Leptospira in a Biphasic Culture Medium Containing Charcoal

A comparison was made of the survival of 28 leptospiral serotypes in Fletcher semisolid medium and in the same medium containing a basal layer of Fletcher medium plus 0.7% of agar and 0.5% of activated animal charcoal. A year after culture, more motile leptospires were observed by microscope examination in the biphasic medium. Two years after culture, 4 serotypes grown in the biphasic medium and 11 in Fletcher medium did not show motile cells. Nineteen of the serotypes maintained in Fletcher medium and 25 in the biphasic medium for 2 years grew on subculture into Fletcher medium. Subcultures from the biphasic medium showed the characteristic leptospiral ring growth earlier during the incubation period.     

Extensin Secreted into the Culture Medium of Tobacco Cells II. Extensin Subfamilies of Similar Composition

Combining acetic acid extraction and high-performance gel chromatographyin guanidine HCl, extensin secreted into the medium by tobacco(Nicotiana tabacum L. var Xanthi) culture cells was separatedinto three component molecules, namely a major 74-kDa, and twominor 45-and 28-kDa components, in addition to larger oligomers.The sizes of these native extensin molecules were first reasonablyassessed using this gel-chromatography system. After deglycosylationwith hydrogen fluoride, the separation was improved and theestimated molecular sizes were reduced to 52 kDa, 34 kDa and18 kDa, respectively. The amino acid compositions of these componentswere similar, and N-terminal sequences of the 52- and 34-kDacomponents coincided. The relative abundance of the componentswas as follows: oligomers, 46%; 52-kDa, 44%; 34-kDa, 7.7%; 18-kDa,2.2%; respectively, on a protein basis (w/w). Fluorography ofthe acid extract of microsomes from cells labelled with ¹⁴C-prolinerevealed only one precursor band of 110-kDa or 42-kDa underthe glycosylating or non-glycosylating conditions, respectively.The smaller components in the medium may be derived, by proteolyticcleavage, from the major extensin molecule after secretion. (Received October 15, 1990; Accepted May 15, 1991)     

栅藻对水环境中镍的累积效应与机理分析

对不同 Scenedesmus 品种的藻细胞从含镍水溶液(10mg/L)中累积金属镍的能力进行了分析,结果表明:藻细胞对镍的生物累积量表现出明显的品种差异性. Scenedesmus quadricauda FACHB 44 和 Scenedesmus quadricauda FACHB 506 表现出很强的累积能力(累积量达到5～6 mg Ni /g干重),而 Scenedesmus sp. FACHB 416 和 Scenedesmus sp. FACHB 489在相同条件对金属镍累积量要少得多(1～1.5 mg Ni /g干重).这种差异可能与不同品种藻细胞间的形态结构和生理特性是相关的.对 S. quadricauda FACHB 44 重金属抗性和累积能力进一步的分析表明, S. quadricauda FACHB 44用于含镍重金属废水处理是非常有效的,在高浓度(100 mg/L)的镍溶液中,藻细胞的最大累积量能达到(26.7 mg Ni/g 干重).对该藻细胞镍累积动力学分析发现:藻细胞对镍的生物累积包括一个快速的被动吸附过程(5 min, 结合70%的镍)和一个缓慢的耗能累积过程(2～3 h时间内的累积量占总量的20%～30%).与其他藻类相比, S. quadricauda FACHB 44对水溶液中镍的耗能累积量明显高于其他藻类.透射电子显微镜(TEM)和X射线能谱(EDX)分析结果均表明,藻细胞耗能累积的镍主要集中在原生质体中,尤以淀粉粒和染色质中为多.     

栅藻对水环境中镍的累积效应与机理分析

对不同Scenedesmus品种的藻细胞从含镍水溶液 (10mg/L)中累积金属镍的能力进行了分析 ,结果表明 :藻细胞对镍的生物累积量表现出明显的品种差异性。ScenedesmusquadricaudaFACHB 4 4和ScenedesmusquadricaudaFACHB 5 0 6表现出很强的累积能力 (累积量达到 5～ 6mgNi /g干重 ) ,而Scenedesmussp .FACHB 4 16和Scenedesmussp .FACHB 4 89在相同条件对金属镍累积量要少得多 (1～ 1.5mgNi /g干重 )。这种差异可能与不同品种藻细胞间的形态结构和生理特性是相关的。对S .quadricaudaFACHB 4 4重金属抗性和累积能力进一步的分析表明 ,S .quadricaudaFACHB 4 4用于含镍重金属废水处理是非常有效的 ,在高浓度 (10 0mg/L)的镍溶液中 ,藻细胞的最大累积量能达到 (2 6 .7mgNi/g干重 )。对该藻细胞镍累积动力学分析发现 :藻细胞对镍的生物累积包括一个快速的被动吸附过程 (5min ,结合 70 %的镍 )和一个缓慢的耗能累积过程 (2～ 3h时间内的累积量占总量的 2 0 %～ 30 % )。与其他藻类相比 ,S .quadricaudaFACHB 4 4对水溶液中镍的耗能累积量明显高于其他藻类。透射电子显微镜(TEM)和X射线能谱 (EDX)分析结果均表明 ,藻细胞耗能累积的镍主要集中在原生质体中 ,尤以淀粉粒和染色质中为多。     

鼎湖山森林土壤渗透水酸度和无机氮含量对模拟氮沉降增加的早期响应

在林地分别喷加0、50、100和150kg N hm^-1a^-1,研究鼎湖山马尾松林、马尾松针阔混交林和季风常绿阔叶林中土壤20cm深渗透水酸度和无机氮含量在开始9个月的变化。结果表明,3种森林对照样方土壤渗透水pH值为3．82—4．24,外加氮处理使其平均降低了0．08—0．18。3个森林对照样方土壤渗透水无机氮平均含量分别为6．14、6．66和11．64mg L^-1,铵态氮占15．0％、11．9％和3．0％。外加氮处理使3种森林土壤渗透水铵态氮和硝态氮含量均有不同程度的提高,这表明外加氮处理不但增加了无机氮从森林土壤流失的潜力,而且使土壤进一步酸化。     

一个快速测定悬浮培养植物细胞生物量的方法的建立

以悬浮培养的玫瑰茄（Hibiscus sabdariffa)细胞为材料,分别探讨了胞外过氧化物酶活力与培养细胞生物量、胞外蛋白浓度与培养细胞生物量之间的相关关系。结果表明,胞外过氧化物酶与培养细胞生物量之间相关性较差,而胞外蛋白浓度与培养细胞生物量之间却具有高度相关性,因此通过胞外蛋白浓度的测量,可以对培养细胞的生物量作出快速的推算。     

Comparative Denitrification of Selected Microorganisms in a Culture Medium and in Autoclaved Soil

The denitrifying behavior of selected soil bacteria was compared in a culture solution and in soil that was sterilized by autoclaving. The essential characteristics concerning nitrate reduction and the formation of nitrogenous gases did not change significantly for most bacteria in the two environments. Bacteria whose denitrification product was nitrous oxide evolved the same gas both in soil and in a liquid system, whereas other bacteria formed only nitrogen gas. The validity of laboratory observations in relation to field studies in the domain of denitrification is discussed and evaluated.     

Marine Sediment Bacteria Harbor Antibiotic Resistance Genes Highly Similar to Those Found in Human Pathogens

The ocean is a natural habitat for antibiotic-producing bacteria, and marine aquaculture introduces antibiotics into the ocean to treat infections and improve aquaculture production. Studies have shown that the ocean is an important reservoir of antibiotic resistance genes. However, there is a lack of understanding and knowledge about the clinical importance of the ocean resistome. We investigated the relationship between the ocean bacterial resistome and pathogenic resistome. We applied high-throughput sequencing and metagenomic analyses to explore the resistance genes in bacterial plasmids from marine sediments. Numerous putative resistance determinants were detected among the resistance genes in the sediment bacteria. We also found that several contigs shared high identity with transposons or plasmids from human pathogens, indicating that the sediment bacteria recently contributed or acquired resistance genes from pathogens. Marine sediment bacteria could play an important role in the global exchange of antibiotic resistance.     

A New Solid Medium for Enumerating Cellulose-Utilizing Bacteria in Soil

A solid medium containing ashed, acid-washed cellulose and a dye, Congo red, has been developed for enumeration of cellulose-utilizing bacteria in soil. Bacteria able to use the cellulose in this medium produced distinct zones of clearing around their colonies. A vivid contrast between the uniform red color of the medium and these halos made this method of differentiation of these organisms superior to other methods.     

Tolerance of Bacteria to High Concentrations of NaCl and Glycerol in the Growth Medium

When compared at similar levels of water activity, glycerol was more inhibitory than sodium chloride to relatively salt-tolerant bacteria and less inhibitory than salt to salt-sensitive species.     

Effect of Ionic Composition of Suspending Solution on Virus Adsorption by a Soil Column

The effect of various electrolytes on the adsorption of poliovirus was measured in 250-cm-long soil columns with ceramic samplers at different depths. Viruses suspended in deionized water moved much farther through the soil than those suspended in tap water, whereas movement in sewage water was intermediate. The salt content of the tap water and sewage water promoted virus adsorption, but evidently the organic compounds in sewage retarded adsorption. When viruses were suspended in chloride solutions of K⁺, Na⁺, Ca⁺, and Mg²⁺, virus adsorption increased as the cation concentration and valence increased. The depth of virus penetration was related to the ionic strength of the solutions. Virus penetration data for NO₃⁻, SO₄²⁻, and H₂PO₄⁻ salts of K⁺, Na⁺, and Ca²⁺ indicated that other anions were more effective than Cl⁻ in promoting virus adsorption. Also, NH₄⁺ was more effective than other cations in limiting the penetration depth of viruses. It seems that ions composed of radicals are more effective than ions composed of single atoms in promoting virus adsorption. Al³⁺ was the most effective ion in limiting virus penetration, probably owing to flocculation of the viruses. Adding AlCl₃ concentrations to secondary sewage effluent to provide an Al³⁺ concentration of 0.1 mM reduced the virus penetration depth to 40 cm. These studies show that the ionic composition of the suspending solutions must be considered in predicting virus penetration depths, and it may be practical to add low concentrations of a flocculating agent such as AlCl₃ to sewage water to limit virus movement through very porous soils.     

Influence of Soil Moisture on Soil Solution Chemistry and Concentrations of Minerals in the Calcicoles Phleum phleoides and Veronica spicata Grown on a Limestone Soil

Veronica spicata and Phleum phleoides are calcicole plants,mainly occurring on neutral or alkaline soil. An experimentof 16 weeks duration was performed in a glasshouse with theobjective of elucidating the influence of soil moisture levelon soil solution chemistry, and biomass concentrations and uptakeof mineral nutrients by the plants. Seven levels of moisture,corresponding to 35–85% of the water holding capacity(WHC) of the soil, were tested. Soil solution HCO₃, P and Mnconcentrations, and pH, increased, whereas Ca, Mg and Zn concentrationsdecreased, with increasing soil moisture. Concentrations ofK were highest at 50–70% WHC. Concentrations and amountsof P, Zn and Mn in the two species were usually related to soilsolution concentrations; these are elements with low solubilityand availability in calcareous soils. Concentrations of nutrientsin biomass were more influenced by soil moisture in V. spicatathan inP. phleoides . This indicates that P. phleoides is morecapable of controlling its uptake of mineral nutrients, whereasV. spicata is sensitive to variations in soil moisture. It isconcluded that variation in soil moisture regime may greatlyinfluence concentrations of mineral nutrients in calcareoussoil solutions and their uptake by plants. Species able to utilizethese solubility fluctuations may have an advantage in competitionfor nutrients. Variation in soil moisture content might evenbe a prerequisite for adequate acquisition of mineral nutrientsand growth of plants on limestone soils, thereby influencingthe field distribution of native plants among habitats. Copyright1999 Annals of Botany Company Calcareous, calcicole, concentration, mineral, moisture, nutrient, Phleum phleoides, soil, soil solution, uptake, Veronica spicata, water.     

Carbonate and Phosphate Precipitation by Saline Soil Bacteria in a Monitored Culture Medium

Carbonate and phosphate precipitation by bacteria isolated from a saline soil was studied in vitro in a liquid culture medium over 45 days. Physicochemical parameters of this medium were continuously monitored using both selective electrodes (continuous monitoring, CM) and individual measurements by other techniques on days 5, 10, 15, 20, 25, 35 and 45 (discontinuous monitoring, DM). In DM, the precipitated minerals were studied (XRD and SEM-EDX) and the saturation index of the mineral phases was analyzed (PHREEQC program). Using the CM and DM data it was possible to distinguish several temporary stages in which both the medium and the mineralogy changed: 1) 0 to 10 days: pH reaches 8.4; significant loss of Mg²⁺ (incorporated into the bacterial biomass) and Ca²⁺ (through mineral precipitation); formation of crystals, although not in sufficient quantity to be studied until day 10. 2) 10 to 25 days: pH decreases but remains above 8; appreciable loss of Mg²⁺ and Ca²⁺ due to formation of spherical carbonate bioliths with traces of phosphates occluded within these carbonates. 3) After 25 days: biomineralization slow down; pH returns to initial values and struvite is formed (idiomorphic prismatic crystals). These trends are in agreement with the findings of other workers, although with some peculiarities regarding stages and types of mineral precipitated. In some cases the struvite contained small quantities of K and Ca, possibly because these are intermediate mineral species between typic-struvite, K-struvite and Ca-struvite. The bacteria-mediated precipitation of carbonates of Ca and/or Mg and phosphates (struvite) by the bacteria from a saline soil is demonstrated. However, struvite was not found in the soils of origin of the bacteria, possibly because it is a metastable mineral in most soils.     

Terpenes Increase the Lipid Dynamics in the Leishmania Plasma Membrane at Concentrations Similar to Their IC50 Values

Although many terpenes have shown antitumor, antibacterial, antifungal, and antiparasitic activity, the mechanism of action is not well established. Electron paramagnetic resonance (EPR) spectroscopy of the spin-labeled 5-doxyl stearic acid revealed remarkable fluidity increases in the plasma membrane of terpene-treated Leishmania amazonensis promastigotes. For an antiproliferative activity assay using 5×10⁶ parasites/mL, the sesquiterpene nerolidol and the monoterpenes (+)-limonene, α-terpineol and 1,8-cineole inhibited the growth of the parasites with IC₅₀ values of 0.008, 0.549, 0.678 and 4.697 mM, respectively. The IC₅₀ values of these terpenes increased as the parasite concentration used in the cytotoxicity assay increased, and this behavior was examined using a theoretical treatment of the experimental data. Cytotoxicity tests with the same parasite concentration as in the EPR experiments revealed a correlation between the IC₅₀ values of the terpenes and the concentrations at which they altered the membrane fluidity. In addition, the terpenes induced small amounts of cell lysis (4–9%) at their respective IC₅₀ values. For assays with high cell concentrations (2×10⁹ parasites/mL), the incorporation of terpene into the cell membrane was very fast, and the IC₅₀ values observed for 24 h and 5 min-incubation periods were not significantly different. Taken together, these results suggest that terpene cytotoxicity is associated with the attack on the plasma membrane of the parasite. The in vitro cytotoxicity of nerolidol was similar to that of miltefosine, and nerolidol has high hydrophobicity; thus, nerolidol might be used in drug delivery systems, such as lipid nanoparticles to treat leishmaniasis.     

不同N、P浓度条件下竹叶眼子菜的生理反应

比较研究了不同氮、磷浓度条件（分别对应于寡营养、中营养、富营养和超营养4种水质状况）培养下竹叶眼子菜的可溶性糖含量、生物量、SOD和POD活性以及ATPase活性变化。在对照（寡营养）中生长的竹叶眼子菜的可溶性糖含量、生物量明显高于其他3组不同氮、磷浓度的处理;试验期间,SOD和ATPase的活性一般表现为先上升再下降的格局,POD则相反 ;SOD活性在第1周内上升,对照条件下活性最小;POD活性在前2周内下降,对照条件下下降幅度最小;叶绿体Mg^2 -ATPase活性在前3周内呈现上升格局,对照条件下表现出较低的活性,线粒体Mg^2 -ATPase活性在前2周出现上升格局,对照条件下该酶活性较低,但其峰值在第3周出现;叶绿体和线粒体Ca^2 -ATPase活性在前2周均呈现上升趋势,超富营养条件下该酶活性最大,且酶活性一般呈现随着氮、磷浓度升高而升高的格局。试验表明竹叶眼子菜比较适合寡营养和中营养水质条件,氮、磷浓度的升高将影响其生理功能,过高的营养条件对竹叶眼子菜是一种胁迫,可抑制其生长。竹叶眼子菜对氮、磷浓度变化虽有一定耐性,但耐性会随时间延长而变弱。     

Selective Culture Medium to Survey the Incidence of Haemophilus Species

A culture medium for the selective isolation of Haemophilus species is described. Bacitracin and nutritional supplements were incorporated in a rich basal agar medium to which rabbit blood was added to distinguish hemolytic species. Colony counts of seven typed strains of H. influenzae on this medium were within practical limits of counts on other media tested for clinical use. The bacitracin medium was as reliable as hemoglobin-agar for detecting H. influenzae and more sensitive for detecting other Haemophilus species in a clinical survey with the advantage of selectivity.