Practical Method for Isolation of Anaerobic Bacteria in the Clinical Laboratory

An inexpensive but practical and simple method is described for providing an environment suitable for the maintenance and survival of clinically significant anaerobic bacteria, consistent with the needs of a large-volume laboratory. This modification of the GasPak procedure is based on two concepts. One involved the storage of "freshly prepared" plating medium under a constant stream of carbon dioxide. The other concept was to use an anaerobe jar, continuously flushed with oxygen-free carbon dioxide, as a "holding receptacle" for the inoculated media Experience with this modification has shown that the number of isolations of anaerobic bacteria from clinical material can be increased significantly.     

An Evaluation of the 'GasKit'Disposable Hydrogen and Carbon Dioxide Generator for the Culture of Anaerobic Bacteria

The new GasKit disposable hydrogen and carbon dioxide generator is a reliable and convenient method for producing anaerobiosis in standard anaerobic jars when compared with a conventional procedure. The generator produces an atmosphere containing 10–12% carbon dioxide by volume which is satisfactory for the culture of exacting anaerobes.     

Effect of Culture Medium and Carbon Dioxide Concentration on Growth of Anaerobic Bacteria Commonly Encountered in Clinical Specimens

Representative strains of anaerobic bacteria from human infections were used to evaluate broth media, gas mixtures, and inocula for use in developing a procedure for performing minimal inhibitory concentration antimicrobic susceptibility tests. Nine commercially available media, including two that were chemically defined, were tested. Tests were performed in atmospheres with carbon dioxide concentrations between 2.5 and 10% and also in the GasPak system (BBL) that had a disposable hydrogen-carbon dioxide generator. Growth curves on each organism grown in schaedler broth and a 5% carbon dioxide atmosphere were used to determine growth characteristics, equate time of the particular growth phases to turbidity readings, and determine the numbers of viable organisms present in the culture. Schaedler broth proved to be most advantageous in combination with an atmosphere of 5% carbon dioxide, 10% hydrogen, and 85% nitrogen. The growth curve studies yielded valuable data on the rapidity and quantity of growth under these conditions. We believe these data have provided information which can be used as the basis for developing a standardized procedure for antimicrobic susceptibility testing for anaerobic bacteria.     

Comparison of recovery of anaerobic bacteria using the Anoxomat, anaerobic chamber, and GasPak jar systems

The Anoxomat system provides an automated evacuation-replacement technique to create an anaerobic or microaerophilic environment in a jar. We evaluated the Anoxomat system for the growth of obligate anaerobes and for the recovery of anaerobic organisms from clinical specimens, and compared its performance to that of an anaerobic chamber and the GasPak System. Of the 54 stock strains tested, the Anoxomat, the chamber, and the GasPak recovered 95%, 95% and 93% at 24 h, respectively. On 29 occasions (51%), the colonies on the Anoxomat plates were slightly larger than those in the chamber and on 17 (30%) occasions larger than the colonies on the GasPak jar plates. At 48 h, the Anoxomat, the chamber, and the GasPak recovered 93.5%, 94.4% and 88.9%, respectively; of 108 anaerobes isolated from 31 clinical specimens. Methylene blue indicators became decolorized (average of 10 tests) within 2 h inside the Anoxomat jars, 2 h 10 min inside the anaerobic chamber, and 2 h 30 min inside the GasPak jars.     

A simple and economical algal culture system for stable isotopic labelling

Summary An alternative method for culturing algae for production of stable isotopically¹³C,¹⁵N-labelled growth media is presented. The culturing principle relies on a closed system connected to a chemical carbon dioxide generator. The system enables economical and labor-inexpensive production of stable isotopically labelled extracts     

Safe Self-contained Carbon Dioxide-Hydrogen Anaerobic System

An anaerobic system is described which allows the microbiologist or hospital technician ease of operation not previously possible with other systems. Gas cylinders, vacuum pumps, valves, and gauges have been eliminated. A new anaerobic lid was developed that is fitted only with a snap-in rubber gasket, a double stainless-steel gauze flash arrestor, and a catalyst holder. The holder contains a palladium pellet catalyst which is active at room temperature and requires no heat for activation. This system was made specifically for use with a disposable hydrogen-carbon dioxide generator and a disposable methylene blue anaerobic indicator. In addition to ease of operation, this jar incorporates unique safety features which eliminate the possibility of a laboratory explosion. An oxygen-free atmosphere composed primarily of nitrogen and carbon dioxide was quickly achieved within the jar to insure maximal growth.     

Gas Exchange of Algae: III. Relation between the Concentration of Carbon Dioxide in the Nutrient Medium and the Oxygen Production of Chlorella pyrenoidosa

The oxygen production of a photosynthetic gas exchanger containing Chlorella pyrenoidosa (1% packed cell volume) was measured when various concentrations of carbon dioxide were present within the culture unit. The internal carbon dioxide concentrations were obtained by manipulating the entrance gas concentration and the flow rate. Carbon dioxide percentages were monitored by means of electrodes placed directly in the nutrient medium. The concentration of carbon dioxide in the nutrient medium which produced maximal photosynthesis was in the range of 1.5 to 2.5% by volume. Results were unaffected by either the level of carbon dioxide in the entrance gas or the rate of gas flow. Entrance gases containing 2% carbon dioxide flowing at 320 ml/min, 3% carbon dioxide at 135 ml/min, and 4% carbon dioxide at 55 ml/min yielded optimal carbon dioxide concentrations in the particular unit studied. By using carbon dioxide electrodes implanted directly in the gas exchanger to optimize the carbon dioxide concentration throughout the culture medium, it should be possible to design more efficient large-scale units.     

Bisource carbon dioxide feeding of the photosynthetic process of microalgae autotrophic cultivation

The mechanism of carbon dioxide saturation of alkaline nutrient medium for the purpose of autotrophic cultivation of microalgae is studied. Carbon dioxide desorption from aqueous solutions of sodium hydrocarbonate is investigated and the influence of chemical equilibria in the solutions on the cultivation of carbon dioxide supply is discussed. It is pointed out that these solutions can be considered to be carriers of the assimilable carbon source for the microalgae cultivation, most efficient at pH 8.5. A comparative estimation of the carbon dioxide and the hydrocarbonic ion as assimilable components of the chemical equilibria under study was done, leading to the conclusion that the more probable component in an alkaline medium autotrophic microalgae cultivation is carbon dioxide. A scheme of bisource feeding of the photosynthetic process, with carbon dioxide for the autotrophic microalgae cultivation in a alkaline nutrient medium, providing a stable and economical cultivation process, is proposed.     

CO2 and the catecholamine content of the adrenal medulla of the rat

In Wistar rats exposed during one hour to mixtures of oxygen and carbon dioxide producing hypoxia, hypercapnia, hyperoxia and hypocapnia, and so on, adrenaline contents of the suprarenals is reduced by high concentration of carbon dioxide (30%), with or without hypoxia. Noradrenaline contents is increased by carbon dioxide (15 to 30%). Hypercapnia is more potent than hypoxia as a suprarenal stimulus.     

配额目标约束下区域减排的最优控制率

研究了区域碳排放配额目标给定下,如何确定一个保障经济平稳增长的最优碳排放控制率问题。提出了一个最优减排率确定的计算流程,并且根据前瞻性原则、人口原则、GDP原则、GDP-人口原则、支付能力原则5种不同的碳排放配额分配原则,确定了中国大部分省市自治区可能的最优减排率。选取5个中国省区上海、山西,湖南、云南以及新疆作为案例地区,在计算得到这些地区的减排率的约束下,分析了在社会福利最大化的情况下各区域分配的配额获得的经济增长路径和能源碳排放路径。模拟结果发现,中国在各种减排约束下各区域的经济增长路径呈阶梯式下降的趋势。能源碳排放呈先升后降的整体趋势,各个省市自治区要实现2030年后不再增加碳排放。得出各东部省份在GDP原则分配的配额约束下减排控制率最小,西部省份在支付能力原则下减排控制率最小,而中部省份则在人口原则下减排控制率较小。5个原则中前瞻性原则适合中国推进的碳减排配额方法。     

Microbial contributions to the evolution of the 'steady state' carbon dioxide system.

Various processes for the production of carbon dioxide by microorganisms are presented. It is postulated that a 'microniche' developed in a reducing environment; a symbiotic relationship between alga-like organisms and bacterium-like organisms in the 'microniche' governed the production of carbon dioxide resulting in the establishment of the steady state carbon dioxide system in the sea.     

Standardized Antimicrobial Disc Susceptibility Testing of Anaerobic Bacteria. I. Susceptibility of Bacteroides fragilis to Tetracycline

A modified Bauer-Kirby-Sherris-Turck method for disc susceptibility testing of anaerobic bacteria is presented. When tetracycline was used against 100 strains of Bacteroides fragilis as a model, reasonably reproducible results were obtained after overnight incubation in both the GasPak atmosphere and an atmosphere achieved by adding 10% CO(2) to a mixture of 10% H(2) and 90% N(2). The minimal inhibitory concentration for the strains determined by the agar dilution technique correlated well with the results of disc tests performed in the GasPak atmosphere with 30-mug tetracycline discs. Among 63 strains isolated from 1970 to the present, only 24 (38.1%) were found to be susceptible to tetracycline.     

Carbon dioxide reversibly inhibits meiosis of Xenopus laevis oocyte and the appearance of the maturation promoting factor

Full-grown Xenopus laevis oocytes were incubated in NaHCO₃ buffer equilibrated with carbon dioxide (5 to 100%). Germinal vesicle breakdown never occurred in spite of the appearance of the characteristic white spot at the animal pole. The effect of carbon dioxide was analyzed during progesterone-induced maturation. Carbon dioxide did not inhibit the early steps of maturation whereas it inhibited germinal vesicle breakdown even when applied 4 hr after the initial hormonal trigger. When oocytes were treated transiently in NaHCO₃ buffer equilibrated with carbon dioxide and further incubated in Tris buffer, drastic delay in the kinetic of germinal vesicle breakdown was observed. Inhibition of progesterone-induced maturation by carbon dioxide treatment is coincident with the time of maturation promoting factor appearance (MPF). On the basis of microinjection experiments of MPF into recipient oocytes, it was also shown that MPF expression is not inhibited by carbon dioxide and thus indicates that the late phase of MPF formation and/or MPF amplification is a carbon dioxide-sensitive period.     

CO2-induced changes in the ERG of the fly,Sarcophaga bullata. A component analysis

The changes in the ERG of the fly, Sarcophaga bullata, with increases in the ambient carbon dioxide concentration are described and analysed using a method of successive algebraic subtraction of normalized responses. On the basis of the behaviour of the five identifiable waves of the ERG before, during, and after carbon dioxide exposure, it is concluded that four components are responsible for the complex ERG. One of these components, which is unaffected by carbon dioxide, apparently arises from the receptor cells, while the other three components have a ganglionic origin. The waveforms of the four proposed components are derived by substracting normalised responses differing with respect to only one of the five waves. The finding that the loss of the waves, through the action of carbon dioxide, proceeds in a sequential and regular manner makes this approach to a component analysis possible. It is suggested that this sequential loss of components does not reflect differences in the basic sensitivity of visual neurons to carbon dioxide, but results from factors such as diffusion barriers and/or chemical processes which establish a decreasing radial concentration gradient of carbon dioxide within the visual ganglia.     

Do fossil plants signal palaeoatmospheric carbon dioxide concentration in the geological past?

Fossil, subfossil, and herbarium leaves have been shown to provide a morphological signal of the atmospheric carbon dioxide environment in which they developed by means of their stomatal density and index. An inverse relationship between stomatal density/index and atmospheric carbon dioxide concentration has been documented for all the studies to date concerning fossil and subfossil material. Furthermore, this relationship has been demonstrated experimentally by growing plants under elevated and reducedcarbon dioxide concentrations. To date, the mechanism that controls the stomatal density response to atmospheric carbon dioxide concentration remains unknown. However, stomatal parameters of fossil plants have been successfully used as a proxy indicator of palaeo-carbon dioxide levels. This paper presents new estimates of palaeo-atmospheric carbon dioxide concentrations for the Middle Eocene (Lutetian), based on the stomatal ratios of fossil Lauraceae species from Bournemouth in England. Estimates of atmospheric carbon dioxide concentrations derived from stomatal data from plants of the Early Devonian, Late Carboniferous, Early Permian and Middle Jurassic ages are reviewed in the light of new data. Semi-quantitative palaeo-carbon dioxide estimates based on the stomatal ratio (a ratio of the stomatal index of a fossil plant to that of a selected nearest living equivalent) have in the past relied on the use of a Carboniferous standard. The application of a new standard based on the present-day carbon dioxide level is reported here for comparison. The resultant ranges of palaeo-carbon dioxide estimates made from standardized fossil stomatal ratio data are in good agreement with both carbon isotopic data from terrestrial and marine sources and long-term carbon cycle modelling estimates for all the time periods studied. These data indicate elevated atmospheric carbon dioxide concentrations during the Early Devonian, Middle Jurassic and Middle Eocene, and reduced concentrations during the Late Carboniferous and Early Permian. Such data are important in demonstrating the long-term responses of plants to changing carbon dioxide concentrations and in contributing to the database needed for general circulation model climatic analogues.     

Changes in the transcutaneous oxygen and carbon dioxide tension during the ischemic test in healthy subjects and under conditions of chronic ischemia of the limb

Changes in the transcutaneous oxygen and carbon dioxide tension during the ischemic test and subsequent reactive hyperemia for 1–2 min in healthy men occur in counterphases. A maximal increase in the carbon dioxide content occurs during the first minute after ischemia. Under conditions of chronic ischemia of the limb, changes in the transcutaneous oxygen and carbon dioxide tension are similar to those in healthy men, but the time intervals are changed and the relative increase in the carbon dioxide content is higher.     

The response of the blood-sucking bug Triatoma infestans to carbon dioxide and other host odours

Behavioural responses of Triatoma infestans larvae to carbon dioxide and other odours of vertebrate origin were investigated in a locomotion compensator. T. infestans oriented towards airstreams enriched with carbon dioxide exhibiting a threshold response between 300 and 400 p.p.m. above the ambient CO(2) background. The accuracy of the oriented response to carbon dioxide improved with stimulus intensity. Remarkably, insects did not show any change in their sensitivity threshold to carbon dioxide with the starvation time. The attractiveness to carbon dioxide depended on the time of the day, i.e. these nocturnal bugs only oriented towards carbon dioxide-loaded airstreams during the first hours of the scotophase. L-lactic acid did not evoke oriented responses when it was presented as a single stimulus in a wide range of intensities. However, a marked synergism was evident when L-lactic acid was combined with a sub-threshold concentration of carbon dioxide. Under this condition, the threshold response to carbon dioxide decreased to 75-150 p.p.m. above ambient CO(2) background. The isomer D-lactic acid evoked no response, either alone or in combination with carbon dioxide. When insects were stimulated with 1-octen-3-ol a significant positive orientation was found. This response was not modified by the addition of carbon dioxide.     

Use of the Minitek system for characterizing lactobacilli.

Incubation of inoculated substrates of the Minitek system in anaerobic GasPak jars provided a method that produced results comparable to those of conventional tubed media for characterizing species of Lactobacillus. The use of sterile mineral oil to overlay inoculated substrate disks was responsible for erroneous results.     

Use of the Minitek system for characterizing lactobacilli.

Incubation of inoculated substrates of the Minitek system in anaerobic GasPak jars provided a method that produced results comparable to those of conventional tubed media for characterizing species of Lactobacillus. The use of sterile mineral oil to overlay inoculated substrate disks was responsible for erroneous results.     

Acclimation to temperature of the response of photosynthesis to increased carbon dioxide concentration in Taraxacum officinale

The relative stimulation of photosynthesis by elevated carbon dioxide in C₃ species normally increases strongly with increasing temperature. This results from the kinetic characteristics of Rubisco, and has potentially important implications for responses of vegetation to increasing atmospheric carbon dioxide. It is often assumed that because Rubisco characteristics are conservative, all C₃ species have the same temperature dependence of the response of photosynthesis to elevated carbon dioxide. However, in this field study of Taraxacum officinale, there were no significant differences in the relative stimulation of photosynthesis by elevated carbon dioxide among days with temperatures ranging from 15 to 34 °C. Nevertheless, short-term measurements indicated a strong temperature dependence of the stimulation. This suggested that acclimation to temperature caused the lack of variation in the seasonal data. Experiments in controlled environments indicated that complete acclimation of the relative stimulation of photosynthesis by elevated carbon dioxide occurred for growth temperatures of 10 – 25 °C. The apparent specificity of Rubisco for carbon dioxide relative to oxygen at 15 °C, as assayed in vivo by measurements of the carbon dioxide concentration at which carboxylation equalled oxygenation, also varied with growth temperature. Changes in the apparent specificity of Rubisco accounted for the acclimation of the temperature dependence of the relative stimulation of photosynthesis by elevated carbon dioxide. It is premature to conclude that low temperatures will necessarily reduce the relative stimulation of photosynthesis caused by rising atmospheric carbon dioxide. This revised version was published online in June 2006 with corrections to the Cover Date.