Effects of resource supplements on mature ciliate biofilms: an empirical test using a new type of flow cell

Biofilm-dwelling consumer communities play an important role in the matter flux of many aquatic ecosystems. Due to their poor accessibility, little is as yet known about the regulation of natural biofilms. Here, a new type of flow cell is presented which facilitates both experimental manipulation and live observation of natural, pre-grown biofilms. These flow cells were used to study the dynamics of mature ciliate biofilms in response to supplementation of planktonic bacteria. The results suggest that enhanced ciliate productivity could be quickly transferred to micrometazoans (ciliate grazers), making the effects on the standing stock of the ciliates detectable only for a short time. Likewise, no effect on ciliates appeared when micrometazoan consumers were ab initio abundant. This indicates the importance of ‘top-down’ control of natural ciliate biofilms. The flow cells used here offer great potential for experimentally testing such control mechanisms within naturally cultivated biofilms.     

Hybridoma perfusion system using a sedimentation device

Summary A novel sedimentation method with a spiral decanter was utilized with a bioreactor for propagation of hybridoma cells at high densities. The live cell concentration was increased and cell lysis was greatly reduced in this system compared to a tangential flow hollow fiber perfusion system. The specific monoclonal antibody productivity was higher than that obtained using a hollow fiber perfusion system or in a batch culture. Cell specific productivity usually declined over time in long term experiments. The use of the sedimentation device eliminated progressive deterioration of reactor performance usually associated with a perfusion device.     

Host Defense Mechanism-Based Rational Design of Live Vaccine

Live attenuated vaccine (LAV), mimicking natural infection, provides an excellent protection against microbial infection. The development of LAV, however, still remains highly empirical and the rational design of clinically useful LAV is scarcely available. Apoptosis and caspase activation are general host antiviral responses in virus-infected cells. Utilizing these tightly regulated host defense mechanisms, we present a novel apoptosis-triggered attenuation of viral virulence as a rational design of live attenuated vaccine with desired levels of safety, efficacy, and productivity. Mutant influenza viruses carrying caspase recognition motifs in viral NP and the interferon-antagonist NS1 proteins were highly attenuated both in vitro and in vivo by caspase-mediated cleavage of those proteins in infected cells. Both viral replication and interferon-resistance were substantially reduced, resulting in a marked attenuation of virulence of the virus. Despite pronounced attenuation, the viruses demonstrated high growth phenotype in embryonated eggs at lower temperature, ensuring its productivity. A single dose vaccination with the mutant virus elicited high levels of systemic and mucosal antibody responses and provided complete protection against both homologous and heterologous lethal challenges in mouse model. While providing a practical means to generate seasonal or pandemic influenza live vaccines, the sensitization of viral proteins to pathogen-triggered apoptotic signals presents a potentially universal, mechanism-based rational design of live vaccines against many viral infections.     

Ecosystem engineers activate mycorrhizal mutualism in salt marshes

Theory predicts that ecosystem engineers should have their most dramatic effects when they enable species, through habitat amelioration, to live in zones where physical and biological conditions would otherwise suppress or limit them. Mutualisms between mycorrhizal fungi and plants are key determinants of productivity and biodiversity in most terrestrial systems, but are thought to be unimportant in wetlands because anoxic sediments exclude fungal symbionts. Our field surveys revealed arbuscular mycorrhizal associations on salt marsh plant roots, but only in the presence of crabs that oxygenate soils as a by-product of burrowing. Field experiments demonstrate that fungal colonization is dependent on crab burrowing and responsible for nearly 35% of plant growth. These results highlight ecosystem engineers as ecological linchpins that can activate and maintain key mutualisms between species. Our findings align salt marshes with other important biogenic habitats whose productivity is reliant on mutualisms between the primary foundation species and micro-organisms.     

Carbon Stocks in Silvopastoral Systems: A Study from Four Communities in Southeastern Ecuador

Agriculture, particularly pasture, is the second largest source of greenhouse gas emissions from tropical regions. Silvopastoral systems may increase carbon pools in pastures while maintaining productivity. Adding trees to pasture provides carbon sinks in woody biomass, and may improve degraded soils and increase the stability of soil carbon pools. In this study we quantified the biomass carbon stored in spontaneous silvopastoral systems in southeastern Ecuador. We compared the stem density, basal area, aboveground biomass, and organic carbon in the top 20 cm of soil in 100 pastures, ranging from 3 to 250 hectares, in four different communities. Aboveground live woody biomass, calculated using allometric equations and two different wood densities, varied from 10.99 to 66.1 Mg per hectare. Soil organic carbon pools ranged from 85.0 to 97.6 Mg per hectare. Stem density, basal area, and aboveground live biomass all positively correlated with pasture age. We found no relationship between pasture age or stem density and soil organic carbon pools. We measured live woody biomass carbon pools of 34–1070 Mg of carbon per farm in these silvopastoral systems. We found no effects on productivity of the herbaceous layer, suggesting that having a low density of trees in pastures could substantially increase the number of trees and the associated carbon sequestration without affecting cattle production.     

Belowground net primary productivity and biomass allocation of a grassland in Inner Mongolia is affected by grazing intensity

The root system of permanent grasslands is of outstanding importance for resource acquisition. Particularly under semi-arid conditions, the acquisition of water and nutrients is highly variable during the vegetation growth period and between years. Additionally, grazing is repeatedly disturbing the functional equilibrium between the root system and the transpiring leaf canopy. However, very few data is available considering grazing effects on belowground net primary productivity (BNPP) and root-shoot dry mass allocation in natural grassland systems. We hypothesise that grazing significantly reduces BNPP due to carbon reallocation to shoot growth. Root biomass and BNPP were estimated by soil coring in 2004, 2005 and 2006 and from ingrowth cores in 2005 and 2006 at one site which has been protected from grazing since 1979 (UG79), at one winter grazing (WG), and one heavily grazed (HG) site. BNPP was estimated from the summation of significant increments of total and live root biomass and from accumulated root biomass of ingrowth cores. Belowground biomass varied from 1,490–2,670 g m^?2 and was significantly lower under heavy grazing than at site UG79. Root turnover varied from 0.23 to 0.33 year^?1 and was not significantly different between sites. Heavy grazing significantly decreased live root biomass and BNPP compared to site UG79. Taking BNPP estimates from live root biomass dynamics and ingrowth cores as the most reliable values, the portion of dry mass allocated belowground relative to total net primary productivity (BNPP/NPP) varied between 0.50–0.66 and was reduced under heavy grazing in 2005, but not in 2006. The positive correlation between cumulative root length density of ingrowth cores and leaf dry matter suggests that the ingrowth core method is suitable for studying BNPP in this semi-arid steppe system. Grazing effects on BNPP and BNPP/NPP should be considered in regional carbon models and estimates of belowground nutrient cycling.     

Understorey productivity in temperate grassy woodland responds to soil water availability but not to elevated [CO2]

Rising atmospheric [CO₂] and associated climate change are expected to modify primary productivity across a range of ecosystems globally. Increasing aridity is predicted to reduce grassland productivity, although rising [CO₂] and associated increases in plant water use efficiency may partially offset the effect of drying on growth. Difficulties arise in predicting the direction and magnitude of future changes in ecosystem productivity, due to limited field experimentation investigating climate and CO₂ interactions. We use repeat near‐surface digital photography to quantify the effects of water availability and experimentally manipulated elevated [CO₂] (eCO₂) on understorey live foliage cover and biomass over three growing seasons in a temperate grassy woodland in south‐eastern Australia. We hypothesised that (i) understorey herbaceous productivity is dependent upon soil water availability, and (ii) that eCO₂ will increase productivity, with greatest stimulation occurring under conditions of low water availability. Soil volumetric water content (VWC) determined foliage cover and growth rates over the length of the growing season (August to March), with low VWC (<0.1 m³ m^?3) reducing productivity. However, eCO₂ did not increase herbaceous cover and biomass over the duration of the experiment, or mitigate the effects of low water availability on understorey growth rates and cover. Our findings suggest that projected increases in aridity in temperate woodlands are likely to lead to reduced understorey productivity, with little scope for eCO₂ to offset these changes.     

Maintaining high anaerobic succinic acid productivity by product removal

During dual-phase fermentations using Escherichia coli engineered for succinic acid production, the productivity and viable cell concentration decrease as the concentration of succinic acid increases. The effects of succinic acid on the fermentation kinetics, yield, and cell viability were investigated by resuspending cells in fresh media after selected fermentation times. The cellular succinic acid productivity could be restored, but cell viability continuously decreased throughout the fermentations by up to 80% and subsequently the volumetric productivity was reduced. Omitting complex nutrients in the resuspension media had no significant effect on cellular succinate productivity and yield, although the viable cell concentration and thus the volumetric productivity was reduced by approximately 20%. By resuspending the cells, the amount of succinate produced during a 100-h fermentation was increased by more than 60%. The results demonstrate that by product removal succinic acid productivity can be maintained at high levels for extended periods of time.     

Improved productivity by reduction of the content of light-harvesting pigment in Chlamydomonas perigranulata

The effects on productivity of the light-harvesting pigment in cells of themarine Chlamydomonas perigranulata were examined using thewild type and a mutant with a lower level of the light-harvesting pigment(LHC-1). We confirmed the photosynthetic characteristics of the wild typeand LHC-1, and these were retained generally in the continuous culturesused for evaluating productivity. The maximum productivity was 1.5 timeshigher in LHC-1 than that in the wild type. The photosynthetic productivityefficiency was higher in LHC-1 than in the wild type. These resultsindicate that the improvement in productivity can be attributed to theimproved photosynthetic productivity efficiency. We conclude that thetechnique of reducing the content of light-harvesting pigment is not onlyavailable for blue-green microalgae, but also for green microalgae.     

Forcing a connection: impacts of single-molecule force spectroscopy on in vivo tension sensing

Mechanical tension plays a large role in cell development ranging from morphology to gene expression. On the molecular level, the effects of tension can be seen in the dynamic arrangement of membrane proteins as well as the recruitment and activation of intracellular proteins. Forces applied to biopolymers during in vitro force measurements offer greater understanding of the effects of tension on molecules in live cells, and experimental techniques involving test tubes and live cells can often overlap. Indeed, when forces exerted on cellular components can be calibrated ex vivo with force spectroscopy, a powerful tool is available for researchers in probing cellular mechanotransduction on the molecular scale. This review will discuss the techniques used in measuring both cellular traction forces and single-molecule force spectroscopy. Emphasis will be placed on the use of fluorescence reporter systems for the development of in vivo tension sensors that can be used for calibration with single molecule force methods.     

杂交瘤细胞培养中摄氧速率(OUR)的在线检测及其与细胞生长及代谢的关系

在批式及灌流培养条件下研究了杂交瘤细胞在无血清培养基中的生长、代谢情况与氧消耗的关系。应用动力学方法在线进行OUR的检测,同时离线取样检测其他参数。结果发现OUR与谷氨酰胺的消耗、抗体的生成及活细胞密度间有明显的相关关系,进一步的分析还发现在对数生长期,OUR与活细胞密度间具有良好的线性关系,qOUR(0.103±0.028)×10^-12mol/cell/h,可以通过它来进行细胞密度的在线检测。并通过以ΔOUR=0时刻作为灌流调整点进行连续灌流培养的初步实验验证了OUR作为培养过程反馈控制参数的可能性。     

Biophysical and economic water productivity of dual-purpose cattle farming

This study analyzes key factors influencing water productivity in cattle rearing, particularly in contexts characterized by water scarcity. This was done through year-round monitoring of on-farm practices within five smallholder farms located in the Saïss area (northern Morocco). The on-farm monitoring protocol consisted of characterizing: (i) volumes of water used for fodder production and distinguished by source (rainfall, surface irrigation and groundwater), (ii) virtual water contained in off-farm feed resources, (iii) total forage biomass production, (iv) dietary rations fed to lactating cows and their calves and (v) milk output and live weight gain. Findings reveal a mean water footprint of 1.62±0.81 and 8.44±1.09 m³/kg of milk and of live weight gain, respectively. Groundwater represented only 13.1% and 2.2% of the total water used to get milk and live weight gain, respectively, while rainfall represented 53.0% and 48.1% of the total water for milk and live weight gain, respectively. The remaining water volumes used came from surface irrigation water (7.4% for milk and 4.0% for live weight gain) and from virtual water (26.5% for milk and 44.7% for live weight gain). The results also revealed a relatively small gross margin per m³ of water used by the herd, not exceeding an average value of US $ 0.05, when considering both milk and live weight. Given the large variability in farm performances, which affect water productivity in cattle rearing throughout the production process, we highlight the potential for introducing a series of interventions that are aimed at saving water, while concurrently improving efficiency in milk production and live weight gain. These interventions should target the chain of production functions that are implemented throughout the process of water productivity in cattle rearing. Moreover, these interventions are of particular importance given our findings that livestock production depends largely upon rainfall, rather than groundwater, in an area afflicted with sustained droughts, overexploitation of groundwater resources and growing water scarcity.     

Effects of cysteine,asparagine, or glutamine limitations in Chinese hamster ovary cell batch and fed-batch cultures

Amino acid availability is a key factor that can be controlled to optimize the productivity of fed-batch cultures. To study amino acid limitation effects, a serum-free chemically defined basal medium was formulated to exclude the amino acids that became depleted in batch culture. The effect of limiting glutamine, asparagine, and cysteine on the cell growth, metabolism, antibody productivity, and product glycosylation was investigated in three Chinese hamster ovary (CHO) cell lines (CHO-DXB11, CHO-K1SV, and CHO-S). Cysteine limitation was detrimental to both cell proliferation and productivity for all three CHO cell lines. Glutamine limitation reduced growth but not cell specific productivity, whereas asparagine limitation had no significant effect on either growth or cell specific productivity. Neither glutamine nor asparagine limitation significantly affected antibody glycosylation. Replenishing the CHO-DXB11 culture with cysteine after 1 day of cysteine limitation allowed the cells to partially recover their growth and productivity. This recovery was not observed after 2 days of cysteine limitation. Based on these findings, a fed-batch protocol was developed using single or mixed amino acid supplementation. Although cell density and antibody concentration were lower compared to a commercial feed, the feeds based on cysteine supplementation yielded comparable cell specific productivity. Overall, this study showed that different amino acid limitations have varied effects on the performance of CHO cell cultures and that maintaining cysteine availability is a critical process parameter for the three cell lines investigated.     

Relationships between rabbit semen characteristics and fertilising ability after insemination

This study aimed to analyse the relationship between rabbit semen characteristics and semen fertilising ability after insemination, which is generally found to be weak. Our hypothesis was that using high semen dilutions (1 : 19), non-oestrus-stimulated does, and homospermic inseminations would make it easier to predict semen fertilising ability. Semen characteristics were evaluated on 275 ejaculates of 128 INRA1001 bucks, distributed into five successive batches. A total of 1970 inseminations were performed. The continuous semen variables were subdivided into three classes of similar size to account for any non-linear relationship between semen characteristics and fertilising ability. Mass motility was divided into two classes according to the presence or absence of waves under microscope observation. Libido, the presence or absence of gel, volume, percentage of progressive sperms, curvilinear velocity, beat frequency of the flagellum, and straightness and linearity of sperm movement did not affect fertility, prolificacy or productivity. It was confirmed that mass motility, estimated by visual observation under the microscope, significantly influenced fertility as well as the percentage of motile and of rapid sperms, and the amplitude of lateral head displacement, estimated by a computer-assisted semen analysis system. To a lesser extent, the percentage of motile cells and of rapid cells significantly influenced prolificacy. Consequently, mass motility and the percentage of motile cells significantly influenced rabbit doe productivity (+1 live births/AI when the semen showed at least a beginning of wave movement, or when the percentage of motile cells was >84%). Interestingly, a gain of 1.5 rabbits was observed when the percentage of rapid cells changed from 64% to 79%, whereas productivity significantly dropped beyond 83% of rapid cells, reflecting a non-linear relationship.     

Biodiversity Increases the Productivity and Stability of Phytoplankton Communities

Global biodiversity losses provide an immediate impetus to elucidate the relationships between biodiversity, productivity and stability. In this study, we quantified the effects of species richness and species combination on the productivity and stability of phytoplankton communities subject to predation by a single rotifer species. We also tested one mechanism of the insurance hypothesis: whether large, slow-growing, potentially-defended cells would compensate for the loss of small, fast-growing, poorly-defended cells after predation. There were significant effects of species richness and species combination on the productivity, relative yield, and stability of phytoplankton cultures, but the relative importance of species richness and combination varied with the response variables. Species combination drove patterns of productivity, whereas species richness was more important for stability. Polycultures containing the most productive single species, Dunaliella, were consistently the most productive. Yet, the most species rich cultures were the most stable, having low temporal variability in measures of biomass. Polycultures recovered from short-term negative grazing effects, but this recovery was not due to the compensation of large, slow-growing cells for the loss of small, fast-growing cells. Instead, polyculture recovery was the result of reduced rotifer grazing rates and persisting small species within the polycultures. Therefore, although an insurance effect in polycultures was found, this effect was indirect and unrelated to grazing tolerance. We hypothesize that diverse phytoplankton assemblages interfered with efficient rotifer grazing and that this “interference effect” facilitated the recovery of the most productive species, Dunaliella. In summary, we demonstrate that both species composition and species richness are important in driving patterns of productivity and stability, respectively, and that stability in biodiverse communities can result from an alteration in consumer functioning. Our findings underscore the importance of predator-prey dynamics in determining the relationships between biodiversity, productivity and stability in producer communities.     

Geographic patterns of coral bioerosion: A productivity hypothesis

The numbers of boring bivalves in corals in large museum collections were used to indicate relative bioerosional damage to the corals. The proportion of corals from different locations that contain boring bivalves is highly correlated with global patterns of plankton primary productivity. The densities of five other, non-boring, groups of plantivores associated with the same corals are similarly correlated with productivity.The proportion of corals containing boring bivalves and the number of boring bivalves per coral head can be ranked by region as follows: eastern Pacific > western Atlantic > Indian Ocean > western Pacific. This ranking also corresponds to primary productivity differences.Measurements of the basal, margin of live tissue, and maximum circumstances of the coral heads indicate that western Atlantic massive corals have more of their skeletal surface exposed to borers, i.e. not covered by live tissue, than do Indo-Pacific corals. Consequently, the former also have weaker basal attachments which suggests that they are more likely to be dislodged during storms. The reason why massive corals in the western Atlantic tend to have less of their skeleton covered by live tissue than corals in the rest of the world is unknown.     

GIS支持下陕西省植被生产有机物价值研究

 在地理信息系统的支持下,利用改进的周广胜NPP模型,根据陕西省植被图和陕西省相关的气候资料,充分考虑植被类型和覆盖度,以植被类型为单位估算了陕西省陆地生态系统NPP物质量进行了测定,结果为：陕西省陆地植被总产量为195.339 5 t·hm－2·a－1,单位面积平均产量为7.25 TDM·hm－2·a-1。得到陕西省植被NPP值列表。以第一性生产物质量为基础,采用能量比市场价格法测定了陕西省各种植被类型的生产力价值,结果是：陕西省的27个植被类型总净第一性生产力价值为358.50亿元。单位面积的生产力价值     

Mathematical modeling and analysis of monoclonal antibody production by hybridoma cells

An attempt has been made to mathematically describe and analyze monoclonal antibody (MAB) productivity of hybridoma cells, with particular emphasis on continuous cultures under unsteady-state conditions. A simple and unstructured general kinetic model that takes account of productivity loss during long-term cultivation, cell proliferation, and the effects of nutrients and toxic products is proposed. The model is verified with data of continuous culture from five different cell lines under a wide range of experimental conditions. Analysis of these results showed that for a reliable assessment of effects of different factors and for comparison of kinetic data on MAB production it is important to consider possible loss of MAB productivity, the time dependence of which can be modeled by an exponential function plus a constant term. Variations of nutrient concentration, particularly that of glucose, glutamine, and serum, can significantly alter MAB production under certain conditions. These effects can be described in terms of saturation kinetic and/or noncompetitive inhibition kinetics. (c) 1996 John Wiley & Sons, Inc.     

Biomass Stock and Productivity of Primeval and Production Beech Forests: Greater Canopy Structural Diversity Promotes Productivity

Our knowledge of temperate broadleaf forest ecology is based mostly on the study of production forests, which lack the terminal stage of forest development and have a simpler stand structure than old-growth and primeval forests. How primeval and production forests differ in net primary production (NPP) is not well known. In three primeval and three nearby production forests of European beech (Fagus sylvatica) in the Slovakian Carpathians, we measured aboveground biomass stocks (live and dead), aboveground NPP (ANPP) and parameters characterizing canopy structural diversity (leaf area index and its spatial variation). Our study aims were (1) to explore the role of canopy structural diversity for ANPP and (2) to assess evidence of a productivity decline in the terminal stage. While aboveground live biomass stocks were on average 20% greater in the primeval forests (386 vs. 320 Mg ha^?1; insignificant difference at two sites), deadwood mass stocks were on average four times larger than in the production forests (86 vs. 19 Mg ha^?1). ANPP was similarly high in the primeval and production forests (10.0 vs. 9.9 Mg ha^?1 y^?1) and did not decrease towards the terminal stage. Production models indicate that, in the primeval forests, about 10% of ANPP (ca. 1 Mg ha^?1 y^?1) was generated by effects related to leaf area heterogeneity, evidencing a positive effect of structural diversity on forest productivity, even though species diversity was low. This study helps to better understand the impact of forest management on the productivity and carbon storage in temperate woodlands.     

Increased protein productivity from immobilized recombinant yeast

The Saccharomyces cerevisiae strain Mc16/p520 has an unstable plasmid, p520, which directs production of a wheat alpha-amylase. The effects of immobilizing this microorganism on the plasmid stability and the specific productivity of the secreted alpha-amylase were investigated. Small gelatin beads were used as the support in both fluidized and packed bed configurations, and the yeast cells were attached by covalent cross-linking with glutaraldehyde. These data were then compared to those for nonimmobilized, suspension cells.Plasmid stability was increased for the immobilized cells during continuous culture at dilution rates both above and below washout. Continuous suspension cultures were not stable and rapidly lost the plasmid. Immobilization caused an increase in specific and volumetric productivity during continuous culture, with a packed bed design resulting in the highest specific productivity.