Biodesulfurization: a model system for microbial physiology research

Biological desulfurization (biodesulfurization) of dibenzothiophene (DBT) by the 4S pathway is a model system for an enviromentally benign way to lower the sulfur content of petroleum. Despite a large amount of effort the efficiency of the 4S pathway is still too low for a commercial oil biodesulfurization process, but the 4S pathway could potentially be used now for commercial processes to produce surfactants, antibiotics, polythioesters and other chemicals and for the detoxification of some chemical warfare agents. Proteins containing disulfide bonds are resistant to temperature, pH, and solvents, but the production of disulfide-rich proteins in microbial hosts is challenging. The study of the 4S pathway can provide insights as to how to maximize the production of disulfide-rich proteins. Engineering of the operon encoding the 4S pathway to contain a greater content of methionine and cysteine may be able to link use of DBT as a sole sulfur source to increasing 4S pathway activity by increasing the nutritional demand for sulfur. This strategy could result in the development of biocatalysts suitable for use in an oil biodesulfurization process, but the study of the 4S pathway can also lead to a better understanding of microbial physiology to optimize activity of a mult-step co-factor-requiring pathway, as well as the production of highly stable industrially relevant enzymes for numerous applications.     

Global microbial ecology: biogeography and diversity of Vibrios as a model

An environmental source of cholera was hypothesized as early as the late nineteenth century by Robert Koch, but not proven because of the ability of Vibrio cholera, the causative agent of cholera, to enter a dormant phase between epidemics. Standard bacteriological procedures for isolation of the vibrios from the environmental samples, including water, between epidemics generally were unsuccessful. Vibrio cholera, a marine vibrio requiring salt for growth, enters into a dormant 'viable but non-culturable' stage when conditions are unfavourable for growth and reproduction. The association of V. cholera with plankton, notably copepods, provides evidence for the environmental origin of cholera, as well as an explanation for the sporadic and erratic nature of cholera epidemics. Thus, the association of V. cholera with plankton was established only recently, allowing analysis of epidemic patterns of cholera, especially in those countries where cholera is endemic. The sporadic and erratic nature of cholera epidemics can now be related to climate and climate events, such as El Ni?o. Since zooplankton have been shown to harbour the bacterium and zooplankton blooms follow phytoplankton blooms, remote sensing can be employed to determine the relationship of cases of cholera with chlorophyll, as well as sea surface temperature (SST), ocean height, and turbidity. Cholera occurs seasonally in Bangladesh with two annual peaks in the number of cases occurring each year. From the data obtained and analysed to date, when the height of the ocean is high and sea surface temperature is also elevated, cholera cases are numerous. When the height is low and sea surface temperature is also low, little or no cholera is recorded. From the examination of data for the 1992-1993 cholera epidemic in India, preliminary comparisons of cholera data for Calcutta show a similar relationship between cholera cases, ocean height and SST. In conclusion, from results of studies of SST, phytoplankton and zooplankton, and their relationships to incidence of cholera, correlation of selected climatological factors and incidence of V. cholera appears to be significant, bringing the potential of predicting conditions conducive to cholera outbreaks closer to reality.     

Big questions, small worlds: microbial model systems in ecology

Although many biologists have embraced microbial model systems as tools to address genetic and physiological questions, the explicit use of microbial communities as model systems in ecology has traditionally been more restricted. Here, we highlight recent studies that use laboratory-based microbial model systems to address ecological questions. Such studies have significantly advanced our understanding of processes that have proven difficult to study in field systems, including the genetic and biochemical underpinnings of traits involved in ecological interactions, and the ecological differences driving evolutionary change. It is the simplicity of microbial model systems that makes them such powerful tools for the study of ecology. Such simplicity enables the high degrees of experimental control and replication that are necessary to address many questions that are inaccessible through field observation or experimentation.     

Saccharomyces sensu stricto as a model system for evolution and ecology

Baker's yeast, Saccharomyces cerevisiae, is not only an extensively used model system in genetics and molecular biology, it is an upcoming model for research in ecology and evolution. The available body of knowledge and molecular techniques make yeast ideal for work in areas such as evolutionary and ecological genomics, population genetics, microbial biogeography, community ecology and speciation. As long as ecological information remains scarce for this species, the vast amount of data that is being generated using S. cerevisiae as a model system will remain difficult to interpret in an evolutionary context. Here we review the current knowledge of the evolution and ecology of S. cerevisiae and closely related species in the Saccharomyces sensu stricto group, and suggest future research directions.     

Complex dynamics in a model microbial system

The forced double-Monod model (for a chemostat with a predator, a prey and periodically forced inflowing substrate) displays quasiperiodicity, phase locking, period doubling and chaotic dynamics. Stroboscopic sections reveal circle maps for the quasiperiodic regimes and noninvertible maps of the interval for the chaotic regimes. Criticality in the circle maps sets the stage for chaos in the model. This criticality may arise with an increase in the period or amplitude of forcing.     

Gas chromatographic metabolic profiling: a sensitive tool for functional microbial ecology

Microbial metabolomics, which consists of a non-targeted analysis of the metabolites released from ('exometabolome') or existing in ('endometabolome') a cell has mostly been used to study the metabolism of particular microbes. Metabolomes also represent a picture of microbial activity and we suggest that the exometabolome may also contain pertinent information for studying microbial interaction networks. Gas chromatography coupled to mass spectrometry is the most commonly used technique in metabolomics studies. It allows a wide range of metabolites to be detected but requires the derivatisation of compounds prior to detection. This type of non-targeted analysis can introduce biases to the detection and quantification of the different metabolites, particularly at the extraction and derivatisation steps. The aims of this study, therefore, were to quantify the sources of variability and to test the sensitivity of the GC metabolic profiling approach to small environmental changes such as shifts in temperature. The temperature sensitivity of metabolic profiles was compared with that of catabolic profiles obtained using Biolog microplates. Analytical variability was compared with biological variability by incubating bacterial strains isolated from soil with fructose at 20 degrees C and by replicating each step of the protocol (incubation, extraction and derivatisation). For both the endo- and the exometabolome, more than 70% of the total variability was of biological origin and principal components analysis clearly separated the strains along the first ordination axis. The endometabolome distinguished bacterial strains at the species level only, whereas separation was evident at the species and group level with the exometabolome. Temperature had a significant but differential effect on the metabolite production of the bacterial strains whilst their catabolic profiles remained relatively unaffected. The exometabolome was more sensitive to temperature shifts than the endometabolome, suggesting that this pool may be of interest for studies in environmental functional ecology.     

Bioaerosol biomonitoring: Sampling optimization for molecular microbial ecology

Bioaerosols (or biogenic aerosols) have largely been overlooked by molecular ecologists. However, this is rapidly changing as bioaerosols play key roles in public health, environmental chemistry and the dispersal ecology of microbes. Due to the low environmental concentrations of bioaerosols, collecting sufficient biomass for molecular methods is challenging. Currently, no standardized methods for bioaerosol collection for molecular ecology research exist. Each study requires a process of optimization, which greatly slows the advance of bioaerosol science. Here, we evaluated air filtration and liquid impingement for bioaerosol sampling across a range of environmental conditions. We also investigated the effect of sampling matrices, sample concentration strategies and sampling duration on DNA yield. Air filtration using polycarbonate filters gave the highest recovery, but due to the faster sampling rates possible with impingement, we recommend this method for fine ‐scale temporal/spatial ecological studies. To prevent bias for the recovery of Gram‐positive bacteria, we found that the matrix for impingement should be phosphate‐buffered saline. The optimal method for bioaerosol concentration from the liquid matrix was centrifugation. However, we also present a method using syringe filters for rapid in‐field recovery of bioaerosols from impingement samples, without compromising microbial diversity for high ‐throughput sequencing approaches. Finally, we provide a resource that enables molecular ecologists to select the most appropriate sampling strategy for their specific research question.     

The ecology and genetics of a host shift: microbotryum as a model system

The need to prevent and cure emerging diseases often precludes their continuing study in situ. We present studies on the process of disease emergence by host shifts using the model system of anther-smut disease (Microbotryum violaceum) on the plant genus Silene (Caryophyllaceae). This system has little direct social impact, and it is readily amenable to experimental manipulation. Our microevolutionary studies have focused on the host shift of Microbotryum from Silene alba (=latifolia; white campion) onto Silene vulgaris (bladder campion) in a population in Virginia. Karyotypic variation shows that the host shift is recent and originates from the disease on sympatric S. alba. Analysis of the spatial pattern of disease shows that the host shift has been contingent on the co-occurrence of the two species at a local scale. Cross-inoculation studies show that families of the new host differ greatly in their susceptibility to the pathogen, indicating the potential for rapid evolution of resistance. Disease expression on the new host is frequently abnormal, suggesting that the pathogen is imperfectly adapted to its new host. In experimental populations, disease transmission within populations of the old host is greater than within populations of the new host. However, there is also a high transmission rate of the disease from the new host back to the old host, suggesting a feedback effect that increases disease prevalence in the community as a whole. Continuing studies of these populations are designed to determine whether this new host-pathogen system is likely to be self-sustaining and to quantify evolutionary changes in both the host and the pathogen.     

Linkage of microbial ecology to phenotype: correlation of rumen microbial ecology to cattle's feed efficiency

Linkage of rumen microbial structure to host phenotypical traits may enhance the understanding of host-microbial interactions in livestock species. This study used culture-independent PCR-denaturing gradient gel electrophoresis (PCR-DGGE) to investigate the microbial profiles in the rumen of cattle differing in feed efficiency. The analysis of detectable bacterial PCR-DGGE profiles showed that the profiles generated from efficient steers clustered together and were clearly separated from those obtained from inefficient steers, indicating that specific bacterial groups may only inhabit in efficient steers. In addition, the bacterial profiles were more likely clustered within a certain breed, suggesting that host genetics may play an important role in rumen microbial structure. The correlations between the concentrations of volatile fatty acids and feed efficiency traits were also observed. Significantly higher concentrations of butyrate (P < 0.001) and valerate (P = 0.006) were detected in the efficient steers. Our results revealed potential associations between the detectable rumen microbiota and its fermentation parameters with the feed efficiency of cattle.     

A functional evenness index for microbial ecology

Microbial ecologists attempting to describe community structures through the use of synthetic parameters face enormous difficulties. These stem in part from the necessity of using standard taxonomic reference levels in a field where the species level is poorly defined. This paper presents an attempt to obviate this problem. A “functional evenness” index (E) is defined using information measures; it is based directly on the characteristics of the bacteria, as determined, for example, with the API 20B method. Comparisons of this index with classic structure indices, such as taxonomic evenness (Pielou) or systematic dominance (Hulburt), show that it behaves like an evenness index, while bypassing the taxonomic study required before computation of the classic indices. Its use is illustrated with samples of aerobic heterotrophic bacteria obtained from brackish lagoon sediments. We are grateful to our statistician colleagues, Dr. Yves Lepage from Université de Montréal and Dr. André Plante from Université du Québec à Montréal. Both have contributed important elements to the discussion presented in this Appendix.     

Arabidopsis thaliana and its wild relatives: a model system for ecology and evolution

The postgenomics era will bring many changes to ecology and evolution. Information about genomic sequence and function provides a new foundation for organismal biology. The crucifer Arabidopsis thaliana and its wild relatives will play an important role in this synthesis of genomics and ecology. We discuss the need for model systems in ecology, the biology and relationships of crucifers, and the molecular resources available for these experiments. The scientific potential of this model system is illustrated by several recent studies in plant–insect interactions, developmental plasticity, comparative genomics and molecular evolution.     

Implications of streamlining theory for microbial ecology

Whether a small cell, a small genome or a minimal set of chemical reactions with self-replicating properties, simplicity is beguiling. As Leonardo da Vinci reportedly said, ‘simplicity is the ultimate sophistication''. Two diverging views of simplicity have emerged in accounts of symbiotic and commensal bacteria and cosmopolitan free-living bacteria with small genomes. The small genomes of obligate insect endosymbionts have been attributed to genetic drift caused by small effective population sizes (N_e). In contrast, streamlining theory attributes small cells and genomes to selection for efficient use of nutrients in populations where N_e is large and nutrients limit growth. Regardless of the cause of genome reduction, lost coding potential eventually dictates loss of function. Consequences of reductive evolution in streamlined organisms include atypical patterns of prototrophy and the absence of common regulatory systems, which have been linked to difficulty in culturing these cells. Recent evidence from metagenomics suggests that streamlining is commonplace, may broadly explain the phenomenon of the uncultured microbial majority, and might also explain the highly interdependent (connected) behavior of many microbial ecosystems. Streamlining theory is belied by the observation that many successful bacteria are large cells with complex genomes. To fully appreciate streamlining, we must look to the life histories and adaptive strategies of cells, which impose minimum requirements for complexity that vary with niche.     

Tribolium beetles as a model system in evolution and ecology

Flour beetles of the genus Tribolium have been utilised as informative study systems for over a century and contributed to major advances across many fields. This review serves to highlight the significant historical contribution that Tribolium study systems have made to the fields of ecology and evolution, and to promote their use as contemporary research models. We review the broad range of studies employing Tribolium to make significant advances in ecology and evolution. We show that research using Tribolium beetles has contributed a substantial amount to evolutionary and ecological understanding, especially in the fields of population dynamics, reproduction and sexual selection, population and quantitative genetics, and behaviour, physiology and life history. We propose a number of future research opportunities using Tribolium, with particular focus on how their amenability to forward and reverse genetic manipulation may provide a valuable complement to other insect models.Subject terms: Model invertebrates, Evolution, Ecology     

Microorganisms with a taste for vanilla: microbial ecology of traditional Indonesian vanilla curing

The microbial ecology of traditional postharvesting processing of vanilla beans (curing) was examined using a polyphasic approach consisting of conventional cultivation, substrate utilization-based and molecular identification of isolates, and cultivation-independent community profiling by 16S ribosomal DNA based PCR-denaturing gradient gel electrophoresis. At two different locations, a batch of curing beans was monitored. In both batches a major shift in microbial communities occurred after short-term scalding of the beans in hot water. Fungi and yeast disappeared, although regrowth of fungi occurred in one batch during a period in which process conditions were temporarily not optimal. Conventional plating showed that microbial communities consisting of thermophilic and thermotolerant bacilli (mainly closely related to Bacillus subtilis, B. licheniformis, and B. smithii) developed under the high temperatures (up to 65 degrees C) that were maintained for over a week after scalding. Only small changes in the communities of culturable bacteria occurred after this period. Molecular analysis revealed that a proportion of the microbial communities could not be cultured on conventional agar medium, especially during the high-temperature period. Large differences between both batches were observed in the numbers of microorganisms, in species composition, and in the enzymatic abilities of isolated bacteria. These large differences indicate that the effects of microbial activities on the development of vanilla flavor could be different for each batch of cured vanilla beans.     

废气处理生物滤器微生物生态学研究进展

废气生物处理是一项新兴的气体污染控制技术,已成为当前环境领域的研究热点.本文概述了生物滤器系统(包括生物过滤器系统和生物滴滤器系统)结构及其去除气体污染物的原理,重点阐述了生物滤器微生物的分离、鉴定及其降解特性,微生物丰度、活性与微生物群落结构多样性之间的相关性,运行条件对微生物群落的影响,微生物群落结构时空变化规律,生物膜形成机理和模型等方面的研究进展,并对今后废气处理生物滤器微生物生态学研究方向进行了展望.