X-ray microanalytic method for measurement of dry matter and elemental content of individual bacteria

A method for the determination of dry matter and elemental content of individual bacterial cells is described. The method is based on energy-dispersive X-ray microanalysis in a transmission electron microscope. A theory for area correction of intensity is developed. Escherichia coli in the late exponential phase of growth and early stationary phase (glucose limited) had an average dry matter content of 278 and 154 fg/cell, respectively. Of the elements detected, sodium, magnesium, phosphorus, sulphur, chlorine, potassium, and calcium together made up 15 to 17% of the dry matter content. A phosphorus content of 4.2 to 5.4% of the dry matter was found in these cells. Volume measurements of air-dried cells gave an average of 1.20 to 1.25 micron3. These results emphasize that dry matter content and elemental composition can be measured directly on single cells from complex microbial communities.     

Bacterial dry matter content and biomass estimations.

Approximately 20% dry-matter content appears to be an accepted standard value for bacterial cells. We have found that the dry-matter content of bacteria may be more than twice as high as generally assumed. The main reason for the low estimates seems to be that proper corrections for intercellular water have not been made when estimating the wet weight of the cells. Using three different bacterial strains, we determined a dry-matter content of cells ranging from 31 to 57%, suggesting not only that the accepted standard value is much too low but also that it is far from standard. To convert bacterial biovolume into biomass (carbon content), we suggest that 0.22 g of C cm-3 should be used as a conversion factor.     

Bacterial dry matter content and biomass estimations

Approximately 20% dry-matter content appears to be an accepted standard value for bacterial cells. We have found that the dry-matter content of bacteria may be more than twice as high as generally assumed. The main reason for the low estimates seems to be that proper corrections for intercellular water have not been made when estimating the wet weight of the cells. Using three different bacterial strains, we determined a dry-matter content of cells ranging from 31 to 57%, suggesting not only that the accepted standard value is much too low but also that it is far from standard. To convert bacterial biovolume into biomass (carbon content), we suggest that 0.22 g of C cm-3 should be used as a conversion factor.     

A new method for non-destructive measurement of biomass, growth rates, vertical biomass distribution and dry matter content based on digital image analysis

BACKGROUND AND AIMS: Biomass is an important trait in functional ecology and growth analysis. The typical methods for measuring biomass are destructive. Thus, they do not allow the development of individual plants to be followed and they require many individuals to be cultivated for repeated measurements. Non-destructive methods do not have these limitations. Here, a non-destructive method based on digital image analysis is presented, addressing not only above-ground fresh biomass (FBM) and oven-dried biomass (DBM), but also vertical biomass distribution as well as dry matter content (DMC) and growth rates. METHODS: Scaled digital images of the plants silhouettes were taken for 582 individuals of 27 grass species (Poaceae). Above-ground biomass and DMC were measured using destructive methods. With image analysis software Zeiss KS 300, the projected area and the proportion of greenish pixels were calculated, and generalized linear models (GLMs) were developed with destructively measured parameters as dependent variables and parameters derived from image analysis as independent variables. A bootstrap analysis was performed to assess the number of individuals required for re-calibration of the models. KEY RESULTS: The results of the developed models showed no systematic errors compared with traditionally measured values and explained most of their variance (R(2) > or = 0.85 for all models). The presented models can be directly applied to herbaceous grasses without further calibration. Applying the models to other growth forms might require a re-calibration which can be based on only 10-20 individuals for FBM or DMC and on 40-50 individuals for DBM. CONCLUSIONS: The methods presented are time and cost effective compared with traditional methods, especially if development or growth rates are to be measured repeatedly. Hence, they offer an alternative way of determining biomass, especially as they are non-destructive and address not only FBM and DBM, but also vertical biomass distribution and DMC.     

On the relation between dry matter and volume of bacteria

Dry matter and volumes of 337 individual bacterial cells with volumes in the range 0.01–7m³ from different origins were measured. An allometric relation was established between dry matter and volume, such that smaller bacteria tended to have a higher dry matter to volume ratio than larger bacteria. The results are compared to results from similar work on algae. The implications for the use of conversion factors are discussed.     

Two measurement methods of leaf dry matter content produce similar results in a broad range of species

BACKGROUND AND AIMS: Leaf dry matter content (LDMC) is widely used as an indicator of plant resource use in plant functional trait databases. Two main methods have been proposed to measure LDMC, which basically differ in the rehydration procedure to which leaves are subjected after harvesting. These are the 'complete rehydration' protocol of Garnier et al. (2001, Functional Ecology 15: 688-695) and the 'partial rehydration' protocol of Vendramini et al. (2002, New Phytologist 154: 147-157). METHODS: To test differences in LDMC due to the use of different methods, LDMC was measured on 51 native and cultivated species representing a wide range of plant families and growth forms from central-western Argentina, following the complete rehydration and partial rehydration protocols. KEY RESULTS AND CONCLUSIONS: The LDMC values obtained by both methods were strongly and positively correlated, clearly showing that LDMC is highly conserved between the two procedures. These trends were not altered by the exclusion of plants with non-laminar leaves. Although the complete rehydration method is the safest to measure LDMC, the partial rehydration procedure produces similar results and is faster. It therefore appears as an acceptable option for those situations in which the complete rehydration method cannot be applied. Two notes of caution are given for cases in which different datasets are compared or combined: (1) the discrepancy between the two rehydration protocols is greatest in the case of high-LDMC (succulent or tender) leaves; (2) the results suggest that, when comparing many studies across unrelated datasets, differences in the measurement protocol may be less important than differences among seasons, years and the quality of local habitats.     

Specific leaf area and dry matter content estimate thickness in laminar leaves

BACKGROUND AND AIMS: Leaf thickness plays an important role in leaf and plant functioning, and relates to a species' strategy of resource acquisition and use. As such, it has been widely used for screening purposes in crop science and community ecology. However, since its measurement is not straightforward, a number of estimates have been proposed. Here, the validity of the (SLA x LDMC)(-1) product is tested to estimate leaf thickness, where SLA is the specific leaf area (leaf area/dry mass) and LDMC is the leaf dry matter content (leaf dry mass/fresh mass). SLA and LDMC are two leaf traits that are both more easily measurable and often reported in the literature. METHODS: The relationship between leaf thickness (LT) and (SLA x LDMC)(-1) was tested in two analyses of covariance using 11 datasets (three original and eight published) for a total number of 1039 data points, corresponding to a wide range of growth forms growing in contrasted environments in four continents. KEY RESULTS AND CONCLUSIONS: The overall slope and intercept of the relationship were not significantly different from one and zero, respectively, and the residual standard error was 0.11. Only two of the eight datasets displayed a significant difference in the intercepts, and the only significant difference among the most represented growth forms was for trees. LT can therefore be estimated by (SLA x LDMC)(-1), allowing leaf thickness to be derived from easily and widely measured leaf traits.     

Protein content, dry mass and chemical composition of individual mast cells related to body growth.

Recently developed quantitative microscopical techniques were used to study relations between body growth and protein content as well as dry mass of individual mast cells. Since previous studies had shown an age-related increase of mast cell content of 5-hydroxytryptamine (5-HT) and heparin, these mast cell components were also included in the present study. The cells were obtained from the peritoneal cavity of rats aged 44--269 days (body weights 189--610 g). All studied mast cell parameters showed an increase that was related to the growth of the animals. The dry mass increased 60%, protein 50%, heparin 50% but 5-HT increased as much as 260% during the studied growth period. There was a mutual and linear correlation between all studied mast cell parameters. Population studies, based on large scale measurements of individual mast cells from young and adult rats, were made. These studies showed that histograms of 5-HT content, protein content and dry mass of individual mast cells were skewed with a tail towards higher values and approximately lognormal. On the other hand, the frequency distribution of heparin content of individual mast cells was approximately normal.     

A method using solubility to predict dry matter digestibility of cellulosic materials.

A rapid chemical procedure based on the solubility of a holocellulose sample in a system of dimethyl sulfoxide with paraformaldehyde has been developed to provide a laboratory method for predicting dry matter digestibility of cellulose containing samples. The amount of dry matter solubilized by the chemical procedure was closely correlated with anaerobic, in vitro, rumen fluid digestion and with digestibility as measured by aerobic Cellulomonas, sp. bacteria. The quantity of solvent and dissolving time had little effect on solubility over a wide range. The method is rapid, well suited for various cellulosic materials, and may be carried out with simple equipment and facilities.     

A new method to estimate individual dry weights of rotifers

Body and egg volumes of 12 rotifer species of Lake Constance were calculated from more than 18 000 individually measured animals using geometric approximations. Body (V _A) and egg (V _E) volumes of various rotifer species have an allometric relationship described by: V _E = 1272 · V _A ^0.3379, while the relation V _E/V _A for each species was constant ranging from 0.016 to 0.87. Dry weight of adult rotifers was assumed to be the sum of the egg dry weight plus a corrected weight increment from egg to adult. It was estimated with the formula W _A = W _E · (1 + a · F _A/F _E) where (a) was varied between 0 and 0.1. Egg dry weight content was assumed to be 35 % of fresh weight. The estimates of dry weight to wet weight ratios ranged from 0.57 % (Asplanchna priodonta) to 29.4 % (Kellicottia longispina). Dry weight estimates obtained with this method are comparable to those obtained with direct measurements of dry weight or carbon content.This investigation was supported by the Deutsche Forschungsgemeinschaft within the Special Collaboration Program (SFB) Cycling of Matter in Lake Constance.     

Quantitative method for measurement of aerotolerance of bacteria and its application to oral indigenous anaerobes.

An index which expressed the aerobic to anaerobic potential was made for bacteria with intermediate tolerance for oxygen. One method used for this analysis was measurement of the relative bacterial growth ratio. The other method was based on the pattern of the absorbancy versus depth plot. The index was applied to oral indigenous anaerobes.     

Specific leaf area and leaf dry matter content as alternative predictors of plant strategies

A key element of most recently proposed plant strategy schemes is an axis of resource capture, usage and availability. In the search for a simple, robust plant trait (or traits) that will allow plants to be located on this axis, specific leaf area is one of the leading contenders. Using a large new unpublished database, we examine the variability of specific leaf area and other leaf traits, the relationships between them, and their ability to predict position on the resource use axis. Specific leaf area is found to suffer from a number of drawbacks; it is both very variable between replicates and much influenced by leaf thickness. Leaf dry-matter content (sometimes referred to as tissue density) is much less variable, largely independent of leaf thickness and a better predictor of location on an axis of resource capture, usage and availability. However, it is not clear how useful dry matter content will be outside northwest Europe, and in particular in dry climates with many succulents.     

Leaf and shoot water content and leaf dry matter content of Mediterranean woody species with different post-fire regenerative strategies

BACKGROUND AND AIMS: Post-fire regeneration is a key process in Mediterranean shrubland dynamics, strongly determining the functional properties of the community. In this study, a test is carried out to determine whether there is co-variation between species regenerative types and functional attributes related to water use. METHODS: An analysis was made of the seasonal variations in leaf relative water content (RWC), leaf dry matter content (LDMC), leaf moisture (LM) and live fine fuel moisture (LFFM) in 30 woody species of a coastal shrubland, with different post-fire regenerative strategies (seeding, resprouting or both). KEY RESULTS: RWC results suggest that the studied resprouters have more efficient mechanisms to reduce water losses and maintain water supply between seasons. In contrast, seeders are more drought tolerant. LDMC is higher in resprouters over the course of the year, suggesting a more efficient conservation of nutrients. The weight of the phylogenetic constraint to understand differences between regenerative strategies tends to be important for LDMC, while it is not the case for variables such as RWC. CONCLUSIONS: Groups of species with different post-fire regenerative strategies (seeders and resprouters) have different functional traits related to water use. In addition to the role of phylogenetical constraints, these differences are also likely to be related to the respective life history characteristics. Therefore, the presence and abundance of species with different post-fire regenerative responses influence the functional properties of the communities.     

Seasonal variability of dry matter content and its relationship with shoot growth and nonstructural carbohydrates

This study assesses how different phases of shoot growth underlie seasonal change in leaf and stem dry matter content (LDMC and SDMC, respectively) of 12 woody Mediterranean species. The relationship between LDMC and nonstructural carbohydrate (NSC) concentrations is also explored and the seasonal vs interspecies variability of LDMC compared. LDMC, SDMC and shoot elongation rate (SER) were measured on a monthly basis for a minimum of 12 months. Bud growth rate (BGR) and NSC concentrations were also assessed in several of the study species. LDMC and SDMC decreased during shoot elongation in spring and increased in summer, showing a significant negative correlation with SER, but were unrelated to BGR. Half of the species analysed showed a positive relationship between LDMC and NSC. Seasonal fluctuations of LDMC within species were higher than interspecies differences, and species ranking was significantly affected by the month of sampling, except during winter months. Seasonal changes in LDMC and SDMC are mainly related to shoot elongation phenology, and NSC sink-source relationships between old and growing organs can explain this relationship in some species. Owing to the high seasonal variability in LDMC, it is recommended that samples for comparative purposes should be collected as close to the winter as possible.     

Dielectric measurement of individual microtubules using the electroorientation method

Little is known about the electrostatic/dynamic properties of microtubules, which are considered to underlie their electrostatic interactions with various proteins such as motor proteins, microtubule-associated proteins, and microtubules themselves (lateral association of microtubules). To measure the dielectric properties of microtubules, we developed an experiment system in which the electroorientation of microtubules was observed under a dark-field microscope. Upon application of an alternating electric field (0.5-1.9 x 10(5) V/m, 10 kHz-3 MHz), the microtubules were oriented parallel to the field line in a few seconds because of the dipole moment induced along their long axes. The process of this orientation was analyzed based on a dielectric ellipsoid model, and the conductivity and dielectric constant of each microtubule were calculated. The analyses revealed that the microtubules were highly conductive, which is consistent with the counterion polarization model-counterions bound to highly negatively charged microtubules can move along the long axis, and this mobility might be the origin of the high conductivity. Our experiment system provides a useful tool to quantitatively evaluate the polyelectrolyte nature of microtubules, thus paving the way for future studies aiming to understand the physicochemical mechanism underlying the electrostatic interactions of microtubules with various proteins.     

A novel method for the measurement of elemental selenium produced by bacterial reduction of selenite

The measurement of elemental selenium (Se⁰) is needed to assess the rate and magnitude of bacteria reduction of selenite or selenate. We have developed a spectrophotometric method for the measurement Se⁰ that is rapid and can be employed to measure the quantity of Se⁰ produced by bacterial cultures. This method employs the use of 1 M Na₂S to convert the insoluble elemental selenium to a red-brown solution and with this method there is a direct correlation between concentration of elemental selenium and the absorption at 500 nm. To demonstrate the utility of this assay, we have followed the reduction of selenite to Se⁰ by Moraxella bovis, and by bacterial consortia in soil and water samples.     

不同沙丘生境主要植物比叶面积和叶干物质含量的比较

研究了生长在不同沙丘生境中 (流动沙丘 ,半固定沙丘和固定沙丘 ) 2 0个植物种 (10个 1年生植物种和 10个多年生植物种 )的比叶面积 (SL A)和叶干物质含量 (L DMC)的变化 ,并且分析了各个沙丘生境的土壤养分特征。结果表明 ,各个植物种的平均 SL A和 L DMC在植物种之间差异显著 ;多数在两种或 3种沙丘生境均有分布的植物其 SL A在不同沙丘生境之间差异显著 ,但是仅有 6个植物种的 L DMC在不同沙丘生境之间表现出差异 (p<0 .0 5 )。与许多研究结果类似 ,1年生植物的 SL A显著大于多年生植物的 SL A,而且两者之间 L DMC存在一定的差异。 1年生植物 SL A和 L DMC之间相关性不显著 ,但多年生植物SL A和 L DMC之间呈显著负相关。综合所有 2 0个植物种可以发现 ,SL A增大时 ,L DMC有下降的趋势