Estimating ecotoxicological risk and impact using indigenous aquatic microbial communities

Emphasis has increased on accuracy in predicting the effect that anthropogenic stress has on natural ecosystems. Although toxicity tests low in environmental realism, such as standardized single species procedures, have been useful in providing a certain degree of protection to human health and the environment, the accuracy of such tests for predicting the effects of anthropogenic activities on complex ecosystems is questionable. The use of indigenous communities of microorganisms to assess the hazard of toxicants in aquatic ecosystems has many advantages. Theoretical and practical aspects of microbial community tests are discussed, particularly in related to widely cited problems in the use of multispecies test systems for predicting hazard. Further standardization of testing protocols using microbial colonization dynamics is advocated on the basis of previous studies, which have shown these parameters to be useful in assessing risk and impact of hazardous substances in aquatic ecosystems.     

Assessing the impact of the biological control agent Bacillus thuringiensis on the indigenous microbial community within the pepper plant phyllosphere

Although biological control agents (BCAs) have been used extensively for controlling insects and pathogens of plants, little is known regarding the effects of such agents on the indigenous microbial communities within the plant phyllosphere. We assessed the effect of the BCA Bacillus thuringiensis (Bt) on the microbial communities within the pepper plant phyllosphere using culture-independent methodologies. Phospholipid fatty acid (PLFA) analysis suggested that the bacterial and fungal biomass were not significantly affected following Bt application. However, principal component analysis of PLFA data indicated that Bt did change the phyllosphere microbial community structure significantly. 16S rRNA gene-directed PCR with denaturing gradient gel electrophoresis (DGGE) also suggested a significant change in the phyllosphere bacterial community structure following Bt inoculation. Phylogenetic analysis of excised DGGE bands suggested a change in bacterial phyla; bands from untreated samples predominantly belonged to the Firmicutes, while Gammaproteobacteria abounded in the treated samples.     

Growth of Salmonella on sprouting alfalfa seeds as affected by the inoculum size, native microbial load and Pseudomonas fluorescens 2-79

Aims: To investigate the growth of salmonellae on sprouting alfalfa seeds as affected by the inoculum size, microbial load and Pseudomonas fluorescens 2–79. Methods and Results: Alfalfa seeds pre‐inoculated with ≤10¹–10³ CFU g^?1 of salmonellae and with or without Ps. fluorescens 2–79 were sprouted in glass jars and the population of salmonellae were determined daily for up to 6 days. The population of salmonellae on germinating seeds reached the maximum 2–3 days after sprouting when total bacterial count reached the maximum (10⁹ CFU g^?1). The population of salmonellae on sprouting seeds not treated with Ps. fluorescens 2–79 showed a net increase of 3–4 log units. However, the population of salmonellae on alfalfa seeds treated with Ps. fluorescens 2–79 showed a net increase of only 1–2 log units. Disinfection of seeds with calcium hypochlorite enhanced the growth of salmonellae. Conclusions: Treatment of seeds with Ps. fluorescens 2–79 reduced the growth of salmonellae by 2–3 log units. Significance and Impact of the Study: The potential of Ps. fluorescens 2–79 as a biological agent for use in control of salmonellae on sprouting seeds was demonstrated and warrants further investigation.     

Rapid and sensitive magnetoelastic biosensors for the detection of Salmonella typhimurium in a mixed microbial population

In this article, we report the results of an investigation into the performance of a wireless, magnetoelastic biosensor designed to selectively detect Salmonella typhimurium in a mixed microbial population. The Langmuir-Blodgett (LB) monolayer technique was employed for antibody (specific to Salmonella sp.) immobilization on rectangular shaped strip magnetoelastic sensors (2 x 0.4 x 0.015 mm). Bacterial binding to the antibody on the sensor surface changes the resonance parameters, and these changes were quantified as a shift in the sensor's resonance frequency. Response of the sensors to increasing concentrations (5 x 10(1) to 5 x 10(8) cfu/ml) of S. typhimurium in a mixture of extraneous foodborne pathogens (Escherichia coli O157:H7 and Listeria monocytogenes) was studied. A detection limit of 5 x 10(3) cfu/ml and a sensitivity of 139 Hz/decade were observed for the 2 x 0.4 x 0.015 mm sensors. Binding kinetics studies have shown that the dissociation constant (K(d)) and the binding valencies for water samples spiked with S. typhimurium was 435 cfu/ml and 2.33 respectively. The presence of extraneous microorganisms in the mixture did not produce an appreciable change in the biosensor's dose response behavior.     

Effect of microbial inoculants on the indigenous actinobacterial endophyte population in the roots of wheat as determined by terminal restriction fragment length polymorphism

The effect of single actinobacterial endophyte seed inoculants and a mixed microbial soil inoculant on the indigenous endophytic actinobacterial population in wheat roots was investigated by using the molecular technique terminal restriction fragment length polymorphism (T-RFLP). Wheat was cultivated either from seeds coated with the spores of single pure actinobacterial endophytes of Microbispora sp. strain EN2, Streptomyces sp. strain EN27, and Nocardioides albus EN46 or from untreated seeds sown in soil with and without a commercial mixed microbial soil inoculant. The endophytic actinobacterial population within the roots of 6-week-old wheat plants was assessed by T-RFLP. Colonization of the wheat roots by the inoculated actinobacterial endophytes was detected by T-RFLP, as were 28 to 42 indigenous actinobacterial genera present in the inoculated and uninoculated plants. The presence of the commercial mixed inoculant in the soil reduced the endophytic actinobacterial diversity from 40 genera to 21 genera and reduced the detectable root colonization by approximately half. The results indicate that the addition of a nonadapted microbial inoculum to the soil disrupted the natural actinobacterial endophyte population, reducing diversity and colonization levels. This was in contrast to the addition of a single actinobacterial endophyte to the wheat plant, where the increase in colonization level could be confirmed even though the indigenous endophyte population was not adversely affected.     

The impact of climatic factors and host density on the long-term population dynamics of vole helminths

Summary The seasonal and long-term population dynamics of helminths parasitizing voles suggested that density-dependent factors might be important in the population dynamics of common species, whereas density-independent factors predominate in the regulation of the rare species. To test this, we used single and multiple regression to analyse the effects of climatic factors and host density on populations of six species of vole helminths over 12 years. The data do support the idea of a difference between common and rare species of helminths, but they clearly do not support the above hypothesis. The common helminths Heligmosomum mixtum (Nematoda) and Catenotaenia sp. (Cestoda) responded to changes in temperature sum (>5° C days) and precipitation during summer. The combined effect of climatic factors and host density explained most of the variation in the long-term dynamics of these common species. By contrast, the long-term dynamics of the rare helminths Paranoplocephala kalelai (Cestoda), Mastophorus muris, Capillaria sp. and Syphacia petrusewiczi (Nematoda) were explained less well by weather and host density than those of the common ones. Furthermore, the common and rare helminths differed in some ways in their responses to climatic factors.     

Modeling the microbial food web

Models of the microbial food web have their origin in the debate over the importance of bacteria as an energetic subsidy for higher trophic levels leading to harvestable fisheries. Conceptualization of the microbial food web preceded numerical models by 10–15 years. Pomeroy's work was central to both efforts. Elements necessary for informative and comprehensive models of microbial loops in plankton communities include coupled carbon and nitrogen flows utilizing a size-based approach to structuring and parameterizing the food web. Realistic formulation of nitrogen flows requires recognition that both nitrogenous and nonnitrogenous organic matter are important substrates for bacteria. Nitrogen regeneration driven by simple mass-specific excretion constants seems to overestimate the role of bacteria in the regeneration process. Quantitative assessment of the link-sink question, in which the original loop models are grounded, requires sophisticated analysis of size-based trophic structures. The effects of recycling complicate calculation of the link between bacteria or dissolved organic matter and mesozooplankton, and indirect effects show that the link might be much stronger than simple analyses have suggested. Examples drawn from a series of oceanic mixed layer plankton models are used to illustrate some of these points. Single-size class models related to traditional P-Z-N approaches are incapable of simulating bacterial biomass cycles in some locations (e.g., Bermuda) but appear to be adequate for more strongly seasonal regimes at higher latitudes.     

Modeling the population dynamics of annual plants with seed bank and density dependent effects

A model is proposed for the population dynamics of an annual plant (Sesbania vesicaria) with a seed bank (i.e. in which a proportion of seeds remain dormant for at least one year). A simple linear matrix model is deduced from the life cycle graph. The dominant eigenvalue of the projection matrix is estimated from demographic parameters derived from field studies. The estimated values for population growth rate () indicates that the study population should be experiencing a rapid exponential increase, but this was not the case in our population.The addition of density dependent effects on seedling survivorship and adult fecundity, effects for which field studies provide evidence, considerably improves our model. Depending on the demographic parameters, the model leads to stable equilibrium, oscillations, or chaos. Study of the behaviour of this model in the parameter space shows that the existence of a seed bank allows higher among-year variation of adult fecundity, without leaving the region of demographic stability. Field data obtained over 3 years confirm this prediction.     

Mean annual population density of Collembola and Acari in the soil and litter of three indigenous South Australian forests

A quantitative study of the soil and litter fauna of three South Australian low open forests was made for two years. The soil and vegetation of the sites were similar but the mean annual rainfall of the areas differed; 635, 690 and 1050 mm/y, respectively for the ‘dry’, ‘medium’ and ‘wet’ sites. Monthly samples were taken of litter, and the upper 0–4 cm and lower 4–8 cm soil layers, and the method used to estimate the numbers of active animals is also described. Mean annual population densities of Collembola and Acari were greatest at the medium site (9.5 and 48.6 × 10³/m², respectively) and least at the dry site (4.9 and 35.1 × 10³/m², respectively) and values are discussed in relation to site organic carbon content. Over all the sites, between 11–21, 67–75 and 10–15% of the collembolan populations, and 10–13, 66–72 and 16–21% of the acarine populations inhabited the litter layer and upper and lower soil layers respectively. Isotomidae were the most abundant collembolan family in each level at each site, but the overall proportion of Entomobryidae increased from the dry to wet site. Prostigmata were the most abundant acarine order in each level at two sites. Cryptostigmata were dominant in the litter layer of the wet site, and their overall proportion increased from the dry to wet site. Seasonal density changes were similar on all sites with minima in the summer and maxima in the winter months. The results obtained here combined with those from other published investigations indicate a trend of faunal population change over various ecosystems in Australia. Prostigmatid mites and iso-tomid Collembola dominate at arid sites. As sites become less arid and the amount of soil organic matter increases, the proportion of cryptostigmatid and mesostigmatid mites increases, and other groups of Collembola, particularly Entomobryidae, become more frequent.     

A method for the regulation of microbial population density during continuous culture at high growth rates

A method for the continuous culture of microorganisms is described which employs growth-dependent pH changes to control the rate of addition of fresh medium to a culture vessel. The apparatus (the phauxostat) supports, at constant pH, longterm continuous culture at rates near or at the maximum of which the organisms are capable. The buffering capacity of the inflowing medium determines the steady-state population density of the culture, but the rate of growth is independent of the buffering capacity. The fundamental theory of operation is tested and some basic parameters of growth are estimated using Escherichia coli B growing continuously in media containing glucose, glycerol or DL-lactate.Joseph C. Wilson Scholar.     

Modeling the impact of single-cell stochasticity and size control on the population growth rate in asymmetrically dividing cells

Microbial populations show striking diversity in cell growth morphology and lifecycle; however, our understanding of how these factors influence the growth rate of cell populations remains limited. We use theory and simulations to predict the impact of asymmetric cell division, cell size regulation and single-cell stochasticity on the population growth rate. Our model predicts that coarse-grained noise in the single-cell growth rate λ decreases the population growth rate, as previously seen for symmetrically dividing cells. However, for a given noise in λ we find that dividing asymmetrically can enhance the population growth rate for cells with strong size control (between a “sizer” and an “adder”). To reconcile this finding with the abundance of symmetrically dividing organisms in nature, we propose that additional constraints on cell growth and division must be present which are not included in our model, and we explore the effects of selected extensions thereof. Further, we find that within our model, epigenetically inherited generation times may arise due to size control in asymmetrically dividing cells, providing a possible explanation for recent experimental observations in budding yeast. Taken together, our findings provide insight into the complex effects generated by non-canonical growth morphologies.     

Allometric scaling of maximum population density: a common rule for marine phytoplankton and terrestrial plants

A primary goal of macroecology is to identify principles that apply across varied ecosystems and taxonomic groups. Here we show that the allometric relationship observed between maximum abundance and body size for terrestrial plants can be extended to predict maximum population densities of marine phytoplankton. These results imply that the abundance of primary producers is similarly constrained in terrestrial and marine systems by rates of energy supply as dictated by a common allometric scaling law. They also highlight the existence of general mechanisms linking rates of individual metabolism to emergent properties of ecosystems.     

Influence of the oral administration of indigenous microorganisms on the resistance of mice to Salmonella infection

The influence of the oral administration of killed bifidobacteria, lactobacteria, bacteroids and fusobacteria on the anti- Salmonella resistance of mice, infected orally with S. dublin, was studied. Bifidobacteria and lactobacteria were shown to produce a dose-dependent immunostimulating effect. The oral administration of killed bifidobacteria and lactobacteria led to the enhanced resistance of mice to Salmonella infection. The oral administration of killed bifidobacteria was conductive to the normalization of the intestinal microflora in dysbacteriosis developing in cases of Salmonella infection. Bacteroids and fusobacteria were found to possess no such effect.     

Effects on microbial activity by extraction of indigenous cells from soil slurries

Abstract: Possible effects on the physiological activity and culturability of soil microorganisms by different soil dispersion procedures, and effects on activity caused by extracting bacteria from soil, were investigated. There was no apparent difference in cfu's with dispersion of a silty loam soil and a loamy sand soil with pyrophosphate as compared to dispersion in NaCl. Substrate-induced respiration was reduced in the silty loam soil, and methanol oxidation was reduced in the loamy sand soil with dispersion in pyrophosphate, and the soil pH was irreversibly increased by the treatment. Extracted bacterial fractions had lower numbers of culturable cells as percentage of the total number of bacteria in each fraction, lower respiration rates and no methanol oxidation activity as compared to the soil slurry both before and after extraction. The physiological activity was apparently not affected by the number of cells extracted. This indicates that the increased extraction rate of indigenous soil bacteria obtained by effective disruption of aggregates and detachment of cells from surfaces, only results in increased extraction of cells that have been physiologically changed as a result of the extraction process.     

下层土壤容重对玉米根际土壤微生物数量及微生物量碳、氮的影响

采用微区池栽模拟试验,研究下层(20～40cm 、40～60cm)土壤容重改变后,玉米根际微生物数量、微生物量C(MBC)和微生物量N(MBN)的动态变化规律.结果表明,玉米根际微生物(细菌、放线菌、真菌)数量、MBC和MBN随土层加深和下层土壤容重增加而降低,且相同层次不同容重的处理间差异达显著水平;不同层次土壤根际微生物数量、MBC和MBN既受本土层容重大小影响,也随相邻土层容重增大其降幅增加,且20～40cm土层容重对土壤微生物数量、MBC和MBN的影响远大于40～60cm土层容重.玉米生育期间,三者受下层土壤容重变化和玉米生长发育的双重影响,且玉米的生长发育过程放大了容重对根际微生物数量、MBC和MBN的影响效果.     

The effect of population density on the growth of an animal population

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Contribution from the Department of Fisheries, Kyoto University.

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Contribution from the Entomological Laboratory, Kyoto University, No. 201.