Antibiotic resistant pathogen outbreak investigation: an interdisciplinary module to teach fundamentals of evolutionary biology

The evolution of resistance to antibiotics provides a timely and relevant topic for teaching undergraduate students evolutionary biology. Here, we present a module incorporating modified sequencing data from eight antibiotic resistant pathogen outbreaks in hospital settings with bioinformatics and phylogenetic analyses. This module uses whole genome sequencing data from hospital outbreaks investigated by the Centers for Disease Control and Prevention to provide examples of antibiotic resistance spread. Students work in groups to analyze outbreak data to identify the bacterial species and antibiotic resistance genes, to infer a phylogenetic tree examining relatedness among isolates, and to determine a possible source of the outbreak. Students then compile their results in individual reports and provide recommendations for preventing the further spread of antibiotic resistant organisms. In addition to providing genomic outbreak data, we include a teaching concepts guide discussing three integral components of the module: how evolutionary biology concepts of natural selection and competition impact antibiotic resistance; outbreak investigation information to aid in phylogenetic analysis and creation of recommendations; and instructions for the bioinformatics protocol. Completion of this module provides students an opportunity to think critically about the evolution of resistance, practice bioinformatics techniques, and relate evolutionary biology to current events.     

Teleological reasoning,not acceptance of evolution,impacts students’ ability to learn natural selection

Background

How acceptance of evolution relates to understanding of evolution remains controversial despite decades of research. It even remains unclear whether cultural/attitudinal factors or cognitive factors have a greater impact on student ability to learn evolutionary biology. This study examined the influence of cultural/attitudinal factors (religiosity, acceptance of evolution, and parents’ attitudes towards evolution) and cognitive factors (teleological reasoning and prior understanding of natural selection) on students’ learning of natural selection over a semester-long undergraduate course in evolutionary medicine.

Method

Pre-post course surveys measured cognitive factors, including teleological reasoning and prior understanding of natural selection, and also cultural/attitudinal factors, including acceptance of evolution, parent attitudes towards evolution, and religiosity. We analyzed how these measures influenced increased understanding of natural selection over the semester.

Results

After controlling for other related variables, parent attitude towards evolution and religiosity predicted students’ acceptance of evolution, but did not predict students’ learning gains of natural selection over the semester. Conversely, lower levels of teleological reasoning predicted learning gains in understanding natural selection over the course, but did not predict students’ acceptance of evolution.

Conclusions

Acceptance of evolution did not predict students’ ability to learn natural selection over a semester in an evolutionary medicine course. However, teleological reasoning did impact students’ ability to learn natural selection.

     

Evolutionary Medicine: A Key to Introducing Evolution

Science teachers can use examples and concepts from evolutionary medicine to teach the three concepts central to evolution: common descent, the processes or mechanisms of evolution, and the patterns produced by descent with modification. To integrate medicine into common ancestry, consider how the evolutionary past of our (or any) species affects disease susceptibility. That humans are bipedal has produced substantial changes in our musculoskeletal system, as well as causing problems for childbirth. Mechanisms such as natural selection are well exemplified in evolutionary medicine, as both disease-causing organism and their targets adapt to one another. Teachers often use examples such as antibiotic resistance to teach natural selection: it takes little alteration of the lesson plan to make explicit that evolution is key to understanding the principles involved. Finally, the pattern of evolution can be illustrated through evolutionary medicine because organisms sharing closer ancestry also share greater susceptibility to the same disease-causing organisms. Teaching evolution using examples from evolutionary medicine can make evolution more interesting and relevant to students, and quite probably, more acceptable as a valid science.     

Evolutionary Distributions and Competition by Way of Reaction-Diffusion and by Way of Convolution

Evolution by natural selection is the most ubiquitous and well understood process of evolution. We say distribution instead of the distribution of the density of populations of phenotypes across the values of their adaptive traits. A phenotype refers to an organism that exhibits a set of values of adaptive traits. An adaptive trait is a trait that a phenotype exhibits where the trait is subject to natural selection. Natural selection is a process by which populations of different phenotypes decline at different rates. An evolutionary distribution (ED) encapsulates the dynamics of evolution by natural selection. The main results are: (i) ED are derived by way of PDE of reaction-diffusion type and by way of integro-differential equations. The latter capture mutations through convolution of a kernel with the rate of growth of a population. The kernel controls the size and rate of mutations. (ii) The numerical solution of a logistic-like ED driven by competition corresponds to a bounded traveling wave solution of population models based on the logistic. (iii) Competition leads to increase in diversity of phenotypes on a single ED. Diversity refers to change in the number of local maxima (minima) within the bounds of values of adaptive traits. (iv) The principle of competitive exclusion in the context of evolution depends, smoothly, on the size and rate of mutations. (v) We identify the sensitivity—with respect to survival—of phenotypes to changes in values of adaptive traits to be an important parameter: increase in the value of this parameter results in decrease in evolutionary-based diversity. (vi) Stable ED corresponds to Evolutionary Stable Strategy; the latter refers to the outcome of a game of evolution.     

Mutational reversions during adaptive protein evolution

Adaptation is often regarded as the sequential fixation of individually, intrinsically beneficial mutations. Contrary to this expectation, we find a surprisingly large number of evolutionary trajectories on which natural selection first favors a mutation, then favors its removal, and later still favors its ultimate restoration during the course of antibiotic resistance evolution. The existence of reversion trajectories implies that natural selection may not follow the most parsimonious path separating two alleles, even during adaptation. Altogether, this discovery highlights the unusual and potentially circuitous routes natural selection can follow during adaptation.     

Students’ Mental Models of Evolutionary Causation: Natural Selection and Genetic Drift

In an effort to understand how to improve student learning about evolution, a focus of science education research has been to document and address students?? naive ideas. Less research has investigated how students reason about alternative scientific models that attempt to explain the same phenomenon (e.g., which causal model best accounts for evolutionary change?). Within evolutionary biology, research has yet to explore how non-adaptive factors are situated within students?? conceptual ecologies of evolutionary causation. Do students construct evolutionary explanations that include non-adaptive and adaptive factors? If so, how are non-adaptive factors structured within students?? evolutionary explanations? We used clinical interviews and two paper and pencil instruments (one open-response and one multiple-choice) to investigate the use of non-adaptive and adaptive factors in undergraduate students?? patterns of evolutionary reasoning. After instruction that included non-adaptive causal factors (e.g., genetic drift), we found them to be remarkably uncommon in students?? explanatory models of evolutionary change in both written assessments and clinical interviews. However, consistent with many evolutionary biologists?? explanations, when students used non-adaptive factors they were conceptualized as causal alternatives to selection. Interestingly, use of non-adaptive factors was not associated with greater understanding of natural selection in interviews or written assessments, or with fewer naive ideas of natural selection. Thus, reasoning using non-adaptive factors appears to be a distinct facet of evolutionary thinking. We propose a theoretical framework for an expert?Cnovice continuum of evolutionary reasoning that incorporates both adaptive and non-adaptive factors, and can be used to inform instructional efficacy in evolutionary biology.     

Using Avida-ED for Teaching and Learning About Evolution in Undergraduate Introductory Biology Courses

Evolution is a complex subject that requires knowledge of basic biological concepts and the ability to connect them across multiple scales of time, space, and biological organization. Avida-ED is a digital evolution educational software environment designed for teaching and learning about evolution and the nature of science in undergraduate biology courses. This study describes our backward design approach to developing an instructional activity using Avida-ED for teaching and learning about evolution in a large-enrollment introductory biology course. Using multiple assessment instruments, we measured student knowledge and understanding of key principles of natural selection before and after instruction on evolution (including the Avida-ED activity). Assessment analysis revealed significant post-instruction learning gains, although certain evolutionary principles (most notably those including genetics concepts, such as the genetic origin of variation) remained particularly difficult for students, even after instruction. Students, however, demonstrated a good grasp of the genetic component of the evolutionary process in the context of a problem on Avida-ED. We propose that: (a) deep understanding of evolution requires complex systems thinking skills, such as connecting concepts across multiple levels of biological organization, and (b) well designed use of Avida-ED holds the potential to help learners build a meaningful and transferable understanding of the evolutionary process. An erratum to this article can be found at     

A field guide for teaching evolution in the social sciences

The theory of evolution by natural selection has begun to revolutionize our understanding of perception, cognition, language, social behavior, and cultural practices. Despite the centrality of evolutionary theory to the social sciences, many students, teachers, and even scientists struggle to understand how natural selection works. Our goal is to provide a field guide for social scientists on teaching evolution, based on research in cognitive psychology, developmental psychology, and education. We synthesize what is known about the psychological obstacles to understanding evolution, methods for assessing evolution understanding, and pedagogical strategies for improving evolution understanding. We review what is known about teaching evolution about nonhuman species and then explore implications of these findings for the teaching of evolution about humans. By leveraging our knowledge of how to teach evolution in general, we hope to motivate and equip social scientists to begin teaching evolution in the context of their own field.     

Metabolic constraints on the evolution of antibiotic resistance

Despite our continuous improvement in understanding antibiotic resistance, the interplay between natural selection of resistance mutations and the environment remains unclear. To investigate the role of bacterial metabolism in constraining the evolution of antibiotic resistance, we evolved Escherichia coli growing on glycolytic or gluconeogenic carbon sources to the selective pressure of three different antibiotics. Profiling more than 500 intracellular and extracellular putative metabolites in 190 evolved populations revealed that carbon and energy metabolism strongly constrained the evolutionary trajectories, both in terms of speed and mode of resistance acquisition. To interpret and explore the space of metabolome changes, we developed a novel constraint‐based modeling approach using the concept of shadow prices. This analysis, together with genome resequencing of resistant populations, identified condition‐dependent compensatory mechanisms of antibiotic resistance, such as the shift from respiratory to fermentative metabolism of glucose upon overexpression of efflux pumps. Moreover, metabolome‐based predictions revealed emerging weaknesses in resistant strains, such as the hypersensitivity to fosfomycin of ampicillin‐resistant strains. Overall, resolving metabolic adaptation throughout antibiotic‐driven evolutionary trajectories opens new perspectives in the fight against emerging antibiotic resistance.     

Bacterial defenses against a natural antibiotic promote collateral resilience to clinical antibiotics

Bacterial opportunistic human pathogens frequently exhibit intrinsic antibiotic tolerance and resistance, resulting in infections that can be nearly impossible to eradicate. We asked whether this recalcitrance could be driven by these organisms’ evolutionary history as environmental microbes that engage in chemical warfare. Using Pseudomonas aeruginosa as a model, we demonstrate that the self-produced antibiotic pyocyanin (PYO) activates defenses that confer collateral tolerance specifically to structurally similar synthetic clinical antibiotics. Non-PYO-producing opportunistic pathogens, such as members of the Burkholderia cepacia complex, likewise display elevated antibiotic tolerance when cocultured with PYO-producing strains. Furthermore, by widening the population bottleneck that occurs during antibiotic selection and promoting the establishment of a more diverse range of mutant lineages, PYO increases apparent rates of mutation to antibiotic resistance to a degree that can rival clinically relevant hypermutator strains. Together, these results reveal an overlooked mechanism by which opportunistic pathogens that produce natural toxins can dramatically modulate the efficacy of clinical antibiotics and the evolution of antibiotic resistance, both for themselves and other members of clinically relevant polymicrobial communities.

This study shows that pyocyanin, a toxin secreted by the opportunistic pathogen Pseudomonas aeruginosa, induces defense responses that decrease the efficacy of structurally-similar clinical antibiotics and accelerate the evolution of antibiotic resistance, both in the producer and in other members of clinically-relevant polymicrobial communities.     

An Alternative Approach: Teaching Evolution in a Natural History Museum Through the Topic of Vector-Borne Disease

Museums play a vitally important role in supporting both informal and formal education and are important venues for fostering public understanding of evolution. The Yale Peabody Museum has implemented significant education programs on evolution for many decades, mostly focused on the museum’s extensive collections that represent the past and present tree of life. Twelve years ago, the Peabody began a series of new programs that explored biodiversity and evolution as it relates to human health. Modern evolutionary theory contributes significantly to our understanding of health and disease, and medical topics provide many excellent and relevant examples to explore evolutionary concepts. The Peabody developed a program on vector-borne diseases, specifically Lyme disease and West Nile virus, which have become endemic in the United States. Both of these diseases have complex transmission cycles involving an intricate interplay among the pathogen, host, and vector, each of which is subject to differing evolutionary pressures. Using these stories, the museum explored evolutionary concepts of adaptation (e.g., the evolution of blood feeding), coevolution (e.g., the “arms race” between host and vector), and variation and selection (e.g., antibiotic resistance) among others. The project included a temporary exhibition and the development of curriculum materials for middle and high school teachers and students. The popularity of the exhibit and some formal evaluation of student participants suggested that this educational approach has significant potential to engage wide audiences in evolutionary issues. In addition it demonstrated how natural history museums can incorporate evolution into a broad array of programs.     

Immunity in a variable world

Immune function is likely to be a critical determinant of an organism's fitness, yet most natural animal and plant populations exhibit tremendous genetic variation for immune traits. Accumulating evidence suggests that environmental heterogeneity may retard the long-term efficiency of natural selection and even maintain polymorphism, provided alternative host genotypes are favoured under different environmental conditions. 'Environment' in this context refers to abiotic factors such as ambient temperature or availability of nutrient resources, genetic diversity of pathogens or competing physiological demands on the host. These factors are generally controlled in laboratory experiments measuring immune performance, but variation in them is likely to be very important in the evolution of resistance to infection. Here, we review some of the literature emphasizing the complexity of natural selection on immunity. Our aim is to describe how environmental and genetic heterogeneities, often excluded from experimentation as 'noise', may determine the evolutionary potential of populations or the potential for interacting species to coevolve.     

The Teaching of Photosynthesis in Secondary School: A History of the Science Approach

In this article we present a synthesis of the research affecting pupils' conceptions of photosynthesis and plant nutrition. The main false conceptions of the pupils identi?ed in this literature review are: that green plants ?nd their food in the soil; that water and mineral salts are suf?cient to the growth of a plant; the role of chlorophyll, where the transformation of luminous energy into chemical energy is never evoked; and air as a source of matter, which is never underlined. Secondly, we are going to see that several of these false conceptions have been developed during history. For example, the famous philosopher Aristotle (384–322 BC) thought that plants receive their food from the soil already elaborated. Several centuries later, the physician and chemist Van Helmont (1677–1644) added more precision to Aristotle’s conception while claiming that plants use only water for their growth. Finally, we will see that the analysis of the false theories developed during history will permit in a context of teaching to valorize the false conceptions of the pupils. Indeed, the history of sciences could incite a teacher to valorize his pupils’ false conceptions while considering them as an indication of difficulties that deserve particular pedagogical and didactic tools. The false conceptions constructed by the pupils don’t have to be ignored in a teaching context because they obey particular reasoning rules, sometimes similar to those that once guided some scientific steps. This view, drawing on false conceptions developed during history, gives a dynamic and human picture of the science very distant from the one sometimes carried on by dogmatic teaching.     

Proposal for Teaching Evolutionary Biology: A bridge between research and educational practice

We present quantitative results for the doctoral thesis of the first-named author of this article. The objective was to recommend and test a teaching proposal for core knowledge of evolutionary biology in secondary education. The focus of the study is ‘Problem cores in teaching’. The ‘Weaving evolutionary thinking’ teaching unit, designed for and tested in this study, entailed: selecting basic evolutionary knowledge from the model of evolution through variation and natural selection; designing a coordinated set of activities, teaching resources and evaluation tools; leading a process of teacher training; and building bridges between: the teaching model, research into the teaching of evolutionary biology, our own teaching experience and that of the secondary school teacher who participated in the project. The data collected through questionnaires show significant differences in favour of the experimental group, which means that the proposal may be favourably assessed in the context of Mexican education, in terms of feasibility, relevance and the pedagogical transformation of knowledge.     

Study of the properties of doxorubicin-resistant cells affected by acute leucosis

The stiffness of cell membrane was found to be one of the factors determining resistance of a cell in vitro to antibiotic doxorubicin action. Membranes of surviving cells are negatively charged (?35 – ?30 mV) and have high values of stiffness (2.2–5.1 μРа) at the doxorubicin concentrations in the medium of 1–500 μg/ml. If the drug concentration and exposure time are being increased, only cells with ‘soft’ membrane (0.25–1 μРа) and positive surface potential (15–29 mV) survive. The data obtained have important prognostic value in studying drug resistance of tumour blood cells and can be used as objective markers of efficiency of the antitumor therapy.     

EVOLUTIONARY ECOLOGY OF DATURA STRAMONIUM L. IN CENTRAL MEXICO: NATURAL SELECTION FOR RESISTANCE TO HERBIVOROUS INSECTS

It has been assumed that herbivores constitute a selective agent for the evolution of plant resistance. However, few studies have tested this hypothesis. In this study, we look at the annual weed Datura stramonium for evidence of current natural selection for resistance to herbivorous insects. Paternal half-sib families obtained through controlled crosses were exposed to herbivores under natural conditions. The plants were damaged by two folivorous insects: the tobacco flea beetle Epitrix parvula and the grasshopper Sphenarium purpurascens. Selection was estimated using a multiple-regression analysis of plant size and of damage by the two herbivores on plant fitness measured as fruit production for both individual phenotypes and family breeding values (genetic analysis). Directional phenotypic selection was detected for both larger plant size and lower resistance to the flea beetles, whereas stabilizing phenotypic selection was revealed for resistance to S. purpurascens. However, performing the same analyses on the breeding values of the characters revealed directional and stabilizing selection only for plant size. Thus, no agreement existed between the results of the two types of analyses, nor was there any detectable potential for genetic change in the studied population because of selection on herbivore resistance. The narrow-sense heritability of every trait studied was small (all <0.1) and not different from zero. The potential for evolutionary response to natural selection for higher resistance to herbivores in the studied population of D. stramonium is probably limited by lack of genetic variation. Natural selection acts on phenotypes, and the detection of phenotypic selection on resistance to herbivores confirms their ecological importance in determining plant fitness. However, evolutionary inferences based solely on phenotypic selection analyses must be interpreted with caution.     

Ecology and evolution of antimicrobial resistance in bacterial communities

Accumulating evidence suggests that the response of bacteria to antibiotics is significantly affected by the presence of other interacting microbes. These interactions are not typically accounted for when determining pathogen sensitivity to antibiotics. In this perspective, we argue that resistance and evolutionary responses to antibiotic treatments should not be considered only a trait of an individual bacteria species but also an emergent property of the microbial community in which pathogens are embedded. We outline how interspecies interactions can affect the responses of individual species and communities to antibiotic treatment, and how these responses could affect the strength of selection, potentially changing the trajectory of resistance evolution. Finally, we identify key areas of future research which will allow for a more complete understanding of antibiotic resistance in bacterial communities. We emphasise that acknowledging the ecological context, i.e. the interactions that occur between pathogens and within communities, could help the development of more efficient and effective antibiotic treatments.Subject terms: Microbial ecology, Antibiotics, Bacterial evolution     

The perception of toxic and non-toxic plants by children and adolescents with regard to gender: implications for teaching botany

ABSTRACT

Human cognition is influenced by natural selection which results in better information retention related to survival and faster visual recognition of potential threat. Plants are excellent models for studying human preferences because of the long evolutionary connectedness of humans with plants as food sources, although research in this field is scarce. We created visual detection tasks to investigate human responses to toxic and non-toxic plants using a sample of children (N = 80) and adolescents (N = 80). As predicted, toxic plants were detected significantly sooner than non-toxic plants. Children showed faster plant detection times than adolescents and females were faster in identification of plants than males. There were, however, no differences in toxic plant identification skills with respect to age and gender. These results suggest that plant toxicity, as an example of survival-relevant information, meets with increased attention on the part of humans and needs to be incorporated into teaching botany.     

The Inability of Primary School to Introduce Children to the Theory of Biological Evolution

A great number of research papers in the English literature of science education present difficulties pupils have in understanding natural selection. Studies show that children have essentialist and teleological intuitive ideas when dealing with organisms and that these biases hinder their ability to understand the theory of evolution by natural selection. Consequently, it is interesting to ascertain if and how the school education offered today deals with the problem, i.e., helps the children confront these biases. To that purpose, this study answered the two following research questions: (a) How is biological evolution presented—from the past to the present day—in the official documentation of primary school education, namely the science curricula and the textbooks of Greece? and (b) what are the conceptions held by Greek primary school teachers of the concepts of evolutionary theory and relevant issues that they have to teach? Our research found that not only are the intuitive ideas not “confronted” but they are also “affirmed” in Greek primary education. This phenomenon, as some other international studies have shown, must not be only a Greek one. A drastic change in the content and structure of primary school curricula and the training of educators is necessary in order to improve and facilitate the teaching of biological evolution.     

Bacterial evolution of antibiotic hypersensitivity

The evolution of resistance to a single antibiotic is frequently accompanied by increased resistance to multiple other antimicrobial agents. In sharp contrast, very little is known about the frequency and mechanisms underlying collateral sensitivity. In this case, genetic adaptation under antibiotic stress yields enhanced sensitivity to other antibiotics. Using large‐scale laboratory evolutionary experiments with Escherichia coli, we demonstrate that collateral sensitivity occurs frequently during the evolution of antibiotic resistance. Specifically, populations adapted to aminoglycosides have an especially low fitness in the presence of several other antibiotics. Whole‐genome sequencing of laboratory‐evolved strains revealed multiple mechanisms underlying aminoglycoside resistance, including a reduction in the proton‐motive force (PMF) across the inner membrane. We propose that as a side effect, these mutations diminish the activity of PMF‐dependent major efflux pumps (including the AcrAB transporter), leading to hypersensitivity to several other antibiotics. More generally, our work offers an insight into the mechanisms that drive the evolution of negative trade‐offs under antibiotic selection.