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NLMR and landscapetools: An integrated environment for simulating and modifying neutral landscape models in R

O_LINeutral landscape models (NLMs) simulate landscape patterns based on theoretical distributions and can be used to systematically study the effect of landscape structure on ecological processes. NLMs are commonly used in landscape ecology to enhance the findings of field studies as well as in simulation studies to provide an underlying landscape. However, their creation so far has been limited to software that is platform dependent, does not allow a reproducible workflow or is not embedded in R, the prevailing programming language used by ecologists.\nC_LIO_LIHere, we present two complementary R packages NLMR and land-scapetools, that allow users to generate, manipulate and analyse NLMs in a single environment. They grant the simulation of the widest collection of NLMs found in any single piece of software thus far while allowing for easy manipulation in a self-contained and reproducible workflow. The combination of both packages should stimulate a wider usage of NLMs in landscape ecology. NLMR is a comprehensive collection of algorithms with which to simulate NLMs. landscapetools provides a utility toolbox which facilitates an easy workflow with simulated neutral landscapes and other raster data.\nC_LIO_LIWe show two example applications that illustrate potential use cases for NLMR and landscapetools: First, an agent-based simulation study in which the effect of spatial structure on disease persistence was studied. Here, spatial heterogeneity resulted in more variable disease outcomes compared to the common well-mixed host assumption. The second example shows how increases in spatial scaling can introduce biases in calculated landscape metrics.\nC_LIO_LISimplifying the workflow around handling NLMs should encourage an uptake in the usage of NLMs. NLMR and landscapetools are both generic frameworks that can be used in a variety of applications and are a further step to having a unified simulation environment in R for answering spatial research questions.\nC_LI

ecology

Intraspecific variation and warming have comparable effects on eco-evolutionary dynamics.

Phenotypic or genotypic variation within species affects ecological processes, from populations to ecosystems. However, whether the ecological imprint of intraspecific variation is substantial compared to key environmental drivers, and persistent enough to carry over to next generations is still questioned. Here, we experimentally showed that intraspecific variation manipulated in a freshwater fish (the European minnow, Phoxinus phoxinus) led to ecological and transgenerational carry-over effects that were as strong as those of varying temperature by 2{degrees}C. Specifically, variation in fish body mass, growth rate and activity altered the size and abundance of prey, which ultimately affected traits and survival of the next fish generation. Temperature variation modulated other ecosystem functions (e.g. litter decomposition) that were also associated to transgenerational carry-over effects. Our results demonstrate that shifting genotypes or phenotypes in wild populations can have substantial and persistent consequences on ecosystems with a similar intensity than climatic variation.

ecology

Environmentally mediated reproductive success predicts breeding dispersal decisions in an early successional amphibian

Dispersal is a central mechanism in ecology and evolution. Dispersal evolution is driven by a trade-off between costs and benefits, which is influenced by inter-individual variability and local environmental conditions (context-dependent dispersal). Many studies have investigated how dispersal decisions may be influenced by environmental factors, including density, predation, and interspecific competition. Yet few have attempted to examine how habitat disturbance may affect the dispersal process in spatially structured populations. In early successional species, one might expect individuals to adjust their dispersal decisions based on two main factors that potentially have an influence on reproductive success: patch size and the level of patch disturbance. In this study, we examined how these two factors affect breeding success and dispersal decisions in an early successional amphibian, the yellow-bellied toad (Bombina variegata). To this end, we used capture-recapture data collected on a spatially structured population occupying 28 breeding patches. We took advantage of recent developments in multievent capture-recapture models to detect signs of context-dependent dispersal. The results revealed that the probability of successful reproduction and the number of newly metamorphosed individuals increased with both the size and the proportion of disturbance of a patch. In addition, our results showed that the factors affecting breeding success also influenced breeding dispersal probability. Large patch size negatively influenced emigration probability; in contrast, it positively influenced immigration probability. Equally, higher disturbance (in terms of the proportion of the patchs surface area disturbed each year) had a strong negative influence on emigration probability and slightly positively affected immigration probability. These findings strongly suggest that individuals make context-dependent dispersal decisions, adjusted to maximize future fitness prospects in a patch, allowing them to better cope with rapid changes in environmental conditions resulting from the ecological succession process. This opens new areas of potential research into the role of dispersal in organism specialization along an ecological succession gradient.

ecology

Fundamental and realized feeding niche breadths of sexual and asexual stick-insects

The factors contributing to the maintenance of sex over asexuality in natural populations remain largely unknown. Ecological divergences between lineages with different reproductive modes could help to maintain reproductive polymorphisms, at least transiently, but there is little empirical information on the consequences of asexuality for the evolution of ecological niches. Here, we investigated how niche breadths evolve following transitions from sexual reproduction to asexuality. We estimated and compared the realized feeding niche breadths of five independently derived asexual Timema stick insect species and their sexual relatives. We found that asexual species had a systematically narrower realized niche than sexual species. To investigate how the narrower realized niches of asexual versus sexual species come about, we quantified the breadth of their fundamental niches but found no systematic differences between reproductive modes. The narrow realized niches found in asexuals are therefore likely a consequence of biotic interactions that constrain realized niche size in asexuals more strongly than in sexuals. Interestingly, the fundamental niche was broader in the oldest asexual species compared to its sexual relative. This broad ecological tolerance may help explain how this species has persisted over more than a million years in absence of sex.

ecology

Information-theoretical approach to measuring landscape complexity

ContextQuantitative grouping of similar landscape patterns is an important part of landscape ecology due to the relationship between a pattern and an underlying ecological process. One of the priorities in landscape ecology is a development of the theoretically consistent framework for quantifying, ordering and classifying landscape patterns.\n\nObjectiveTo demonstrate that the Information Theory as applied to a bivariate random variable provides a consistent framework for quantifying, ordering, and classifying landscape patterns.\n\nMethodsAfter presenting Information Theory in the context of landscapes, information-theoretical metrics were calculated for an exemplar set of landscapes embodying all feasible configurations of land cover patterns. Sequences and 2D parametrization of patterns in this set were performed to demonstrate the feasibility of Information Theory for the analysis of landscape patterns.\n\nResultsUniversal classification of landscape into pattern configuration types was achieved by transforming landscapes into a 2D space of weakly correlated information-theoretical metrics. An ordering of landscapes by any single metric cannot produce a sequence of continuously changing patterns. In real-life patterns, diversity induces complexity - increasingly diverse patterns are increasingly complex.\n\nConclusionsInformation theory provides a consistent, theory-based framework for the analysis of landscape patterns. Information-theoretical parametrization of landscapes offers a method for their classification.

ecology

Towards an applied metaecology

The complexity of ecological systems is a major challenge for practitioners and decision-makers who work to avoid, mitigate and manage environmental change. Here, we illustrate how metaecology - the study of spatial interdependencies among ecological systems through fluxes of organisms, energy, and matter - can enhance understanding and improve managing environmental change at multiple spatial scales. We present several case studies illustrating how the framework has leveraged decision-making in conservation, restoration and risk management. Nevertheless, an explicit incorporation of metaecology is still uncommon in the applied ecology literature, and in action guidelines addressing environmental change. This is unfortunate because the many facets of environmental change can be framed as modifying spatial context, connectedness and dominant regulating processes - the defining features of metaecological systems. Narrowing the gap between theory and practice will require incorporating system-specific realism in otherwise predominantly conceptual studies, as well as deliberately studying scenarios of environmental change.

ecology

Eco-evolutionary processes underlying early warning signals of population declines

O_LIEnvironmental change can impact the stability of populations and can cause rapid declines in abundance. Abundance-based warning signals have been proposed to predict such declines, but these have been shown to have limited success, leading to the development of warning signals based on the distribution of fitness-related traits such as body size. C_LIO_LIThe dynamics of such traits in response to external environmental perturbations are controlled by a range of underlying factors such as reproductive rate, genetic variation, and plasticity. However, it remains unknown how such ecological and evolutionary factors affect the stability landscape of populations and the detectability of abundance and trait-based warning signals of population decline. C_LIO_LIHere, we apply a trait-based demographic approach and investigate both trait and population dynamics in response to gradual changes in the environment. We explore a range of ecological and evolutionary constraints under which the stability of a population may be affected. C_LIO_LIWe show both analytically and with model-based simulations that strength of abundance-based early warning signals is significantly affected by ecological and evolutionary factors. C_LIO_LIFinally, we show that a unified approach, combining trait- and abundance-based information, significantly improves our ability to predict population declines. Our study suggests that the inclusion of trait dynamic information alongside generic warning signals should provide more accurate forecasts of the future state of biological systems. C_LI

ecology

Extinction times in diffusive public good population dynamics

The co-evolutionary dynamics of competing populations can be strongly affected by frequency-dependent selection and population structure in space. As co-evolving populations grow into a spatial domain, their initial spatial arrangement, as well as their growth rate differences determine the dynamics. Here, we are interested in the dynamics of producers and free-rider co-evolution in the context of an ecological public good that is produced by a sub-population but evokes growth benefits to all individuals. We consider the spatial growth dynamics in one, two and three dimensions by modeling producer cell, free-rider cell and public good densities in space, driven by birth, death and diffusion. Typically, one population goes extinct. We find that uncorrelated initial spatial structures do not influence the time to extinction in comparison to the well-mixed system. We derive a slow manifold solution in order to estimate the time to extinction of either free-riders or producers. For invading populations, i.e. for populations that are initially highly segregated, we observe a traveling wave, whose speed can be calculated to improve the extinction time estimate by a simple superposition of the two times. Our results show that local effects of spatial dynamics evolve independently of the dynamics of the mean populations. Our considerations provide quantitative predictions for the transient dynamics of cooperative traits under pressure of extinction, and a potential experiment to derive elusive details of the fitness function of an ecological public goods game through extinction time observations.\n\nAuthor SummaryEcological public goods (PG) relationships emerge in growing cellular populations, for example between bacteria and cancer cells. We study the eco-evolutionary dynamics of a PG in populations that grow in space. In our model, public good-producer cells and free-rider cells can grow according to their own birth and death rates. Co-evolution occurs due to public good-driven surplus in the intrinsic growth rates and a cost to producers. A net growth rate benefit to free-riders leads to the well-known tragedy of the commons in which producers go extinct. What is often omitted from discussions is the time scale on which this extinction can occur, especially in spatial populations. We derive analytical estimates of the time to extinction in different spatial settings, and identify spatial scenarios in which extinction takes long enough such that the tragedy of the commons never occurs within the lifetime of the populations. Using numerical simulations we analyze the deviations from analytical predictions. Our results have direct implications for inferring ecological public good game properties from in vitro and in vivo experimental observations.

ecology

Both environmental filtering and intraspecific variation shape small mammals' elementomes

The biogeochemical niche hypothesis (BNH) proposes the multi-elemental composition of organisms - their elementome - as a new ecological dimension. However, which ecological factors shape elementome assembly remains little known, especially in animals. Here, we studied the mandibular elementome of two sympatric small mammals - Apodemus flavicollis and Clethrionomys glareolus - to assess how intraspecific variability (ontogenetic changes in body mass and sex under the vertebrate bone hypothesis; VBH) and environmental filtering (season and habitat) shape essential and non-essential elementome assembly. Species showed moderate elementome segregation and seasonal niche partitioning, with implications for coexistence. Ontogenetic body mass predicted elemental variation and calcium substitution, with several hypermetric scalings in autumn indicating strong departures from mass-invariant homeostasis. Finally, our results suggest a dichotomy: essential elementomes were mainly driven by intraspecific variation, whereas non-essential elementomes were rather shaped by environmental filtering. Our results position animal elementomes as an integrative ecological dimension linking organismal biology, species interactions, and environmental filtering across individuals, populations, and species.

ecology

Developments in the European parasitoid community of Dryocosmus kuriphilus

The invasive gallwasp Dryocosmus kuriphilus was first detected in Italy in 2002, although likely to have initially arrived in the late 90s. Its ability to utilise sweet chestnut species non-native to its original Chinese range has allowed it to spread rapidly, and throughout Europe via European sweet chestnut Castanea sativa. Given the severity of its impact on C. sativa crop production, particularly in Mediterranean countries, previous studies have aimed to assess damage levels caused by D. kuriphilus, the efficacy of and potential non-target effects of the introduced biocontrol agent Torymus sinensis, and the possibility of regulation by native parasitoids. As yet a broad overview and analytical synthesis of these native parasitoid communities are absent. This review focuses on important aspects of the D. kuriphilus invasion. In particular, the invasion history and currently known distribution of D. kuriphilus, and several aspects of its associated parasitoid community. For native species to plausibly suppress D. kuriphilus, we might expect rates of parasitoid attack to increase with establishment time as native populations adapt to exploit the new resource, and this is a key focus of the review. We answer the following questions: 1) What is the distribution of D. kuriphilus in Europe, and has D. kuriphilus fully utilised the available niche space within its 20+ years in Europe? 2) Which species of native parasitoids attack D. kuriphilus in Europe and what are their ecological characteristics? 3) How consistent is the parasitoid community of D. kuriphilus across its range, and are there signs of convergence over time? 4) What effect does establishment time have on the species richness and abundance of parasitoid communities? We report the following: 1) D. kuriphilus has expanded its range throughout Europe and is present in nearly every major region where sweet chestnut is present. Native and non-native naturalised chestnut forests may be less susceptible to invasion than areas of industry due to differing socioeconomic and ecological factors, though areas with large chestnut industries also tend to be in the most heavily forested areas in the non-native range of sweet chestnut. D. kuriphilus has reportedly been eradicated from some countries, and effectively eradicated in a number of countries implementing biocontrol with T. sinensis, although successive invasions from neighbouring regions are still possible, and eradication may be transient. 2) 72 parasitoid species are identified attacking D. kuriphilus in Europe (far more species than any other gallwasp in the Western Palearctic). Its members are predominantly oak gallwasp parasitoids (82% of species), followed by gall-specialists of different host plants, leaf miner parasitoids and a minority of others with differing host life stages and ecologies. Parasitoids attacking D. kuriphilus are dominated by idiobiont ectoparasitoids of the superfamily Chalcidoidea (>96%). 3) The parasitoid community is highly variable, both temporally and spatially, although the vast proportion (>95%) of parasitoids at any one time are composed of locally common generalist oak gall parasitoids. The most common members include Bootanomyia dorsalis, Eupelmus urozonus, Eurytoma brunniventris, Mesopolobus sericeus and Torymus flavipes. 4) The length of establishment time has minimal effect on the species richness, abundance, and composition of the community, suggesting that regulation by natives, if it occurs, may take longer than the 20+ years that D. kuriphilus has persisted. While little evidence of increasing parasitoid attack of D. kuriphilus is apparent, we exercise caution by stating that the heterogeneity in available data are large, and that common biocontrol interventions using T. sinensis interrupt the natural process of community development dramatically. D. kuriphilus has been present in Europe for nearly three decades and few localities have repeated years of data collection. Even fewer studies have communities with establishment times exceeding ten years. Proper biocontrol by natives may not occur within short timeframes, although studies of other gallwasp invaders find similar results over periods exceeding 40 years. Given that many countries have chosen to implement T. sinensis for biocontrol, the focus may be better spent monitoring native gall communities for potential non-target effects.

ecology

Pathways to social evolution: reciprocity, relatedness, and synergy

Many organisms live in populations structured by space and by class, exhibit plastic responses to their social partners, and are subject to non-additive ecological and fitness effects. Social evolution theory has long recognized that all of these factors can lead to different selection pressures but has only recently attempted to synthesize how these factors interact. Using models for both discrete and continuous phenotypes, we show that analyzing these factors in a consistent framework reveals that they interact with one another in ways previously overlooked. Specifically, behavioral responses (reciprocity), genetic relatedness, and synergy interact in non-trivial ways that cannot be easily captured by simple summary indices of assortment. We demonstrate the importance of these interactions by showing how they have been neglected in previous synthetic models of social behavior both within and between species. These interactions also affect the level of behavioral responses that can evolve in the long run; proximate biological mechanisms are evolutionarily stable when they generate enough responsiveness relative to the level of responsiveness that exactly balances the ecological costs and benefits. Given the richness of social behavior across taxa, these interactions should be a boon for empirical research as they are likely crucial for describing the complex relationship linking ecology, demography, and social behavior.

Evolutionary Biology

Biochemical ‘Cambrian’ explosion-implosions: the generation and pruning of genetic codes

Tlusty's topological analysis of the genetic code suggests ecosystem changes in available metabolic free energy that predated the aerobic transition enabled a punctuated sequence of increasingly complex genetic codes and protein translators. These coevolved via a `Cambrian explosion' until, very early on, the ancestor of the present narrow spectrum of protein machineries became evolutionarily locked in at a modest level of fitness reflecting a modest embedding metabolic free energy ecology. Similar biochemical `Cambrian singularities' must have occurred at different scales and levels of organization on Earth, with competition or chance-selected outcomes frozen at a far earlier period than the physical bauplan Cambrian explosion. Other examples might include explosive variations in mechanisms of photosynthesis and subsequent oxygen metabolisms. Intermediate between Cambrian bauplan and genetic code, variants of both remain today, even after evolutionary pruning, often protected in specialized ecological niches. This suggests that, under less energetic astrobiological ecologies, a spectrum of less complicated reproductive codes may also survive in specialized niches.

Evolutionary Biology

Reproductive isolation of hybrid populations driven by genetic incompatibilities

Despite its role in homogenizing populations, hybridization has also been proposed as a means to generate new species. The conceptual basis for this idea is that hybridization can result in novel phenotypes through recombination between the parental genomes, allowing a hybrid population to occupy ecological niches unavailable to parental species. A key feature of these models is that these novel phenotypes ecologically isolate hybrid populations from parental populations, precipitating speciation. Here we present an alternative model of the evolution of reproductive isolation in hybrid populations that occurs as a simple consequence of selection against incompatibilities. Unlike previous models, our model does not require small population sizes, the availability of new niches for hybrids or ecological or sexual selection on hybrid traits. We show that reproductive isolation between hybrids and parents evolves frequently and rapidly under this model, even in the presence of substantial ongoing migration with parental species and strong selection against hybrids. Our model predicts that multiple distinct hybrid species can emerge from replicate hybrid populations formed from the same parental species, potentially generating patterns of species diversity and relatedness that resemble an adaptive radiation.

Evolutionary Biology

BEEtag : a low-cost, image-based tracking system for the study of animal behavior and locomotion

A fundamental challenge common to studies of animal movement, behavior, and ecology is the collection of high-quality datasets on spatial positions of animals as they change through space and time. Recent innovations in tracking technology have allowed researchers to collect large and highly accurate datasets on animal spatiotemporal position while vastly decreasing the time and cost of collecting such data. One technique that is of particular relevance to the study of behavioral ecology involves tracking visual tags that can be uniquely identified in separate images or movie frames. These tags can be located within images that are visually complex, making them particularly well suited for longitudinal studies of animal behavior and movement in naturalistic environments. While several software packages have been developed that use computer vision to identify visual tags, these software packages are either (a) not optimized for identification of single tags, which is generally of the most interest for biologist, or (b) suffer from licensing issues, and therefore their use in the study of animal behavior has been limited. Here, we present BEEtag, an open-source, image-based tracking system in Matlab that allows for unique identification of individual animals or anatomical markers. The primary advantages of this system are that it (a) independently identifies animals or marked points in each frame of a video, limiting error propagation, (b) performs well in images with complex background, and (c) is low-cost. To validate the use of this tracking system in animal behavior, we mark and track individual bumblebees (Bombus impatiens) and recover individual patterns of space use and activity within the hive. Finally, we discuss the advantages and limitations of this software package and its application to the study of animal movement, behavior, and ecology.

Animal Behavior and Cognition

Estimating the effect of competition on trait evolution using maximum likelihood inference

Many classical ecological and evolutionary theoretical frameworks posit that competition between species is an important selective force. For example, in adaptive radiations, resource competition between evolving lineages plays a role in driving phenotypic diversification and exploration of novel ecological space. Nevertheless, current models of trait evolution fit to phylogenies and comparative datasets are not designed to incorporate the effect of competition. The most advanced models in this direction are diversity-dependent models where evolutionary rates depend on lineage diversity. However, these models still treat changes in traits in one branch as independent of the value of traits on other branches, thus ignoring the effect of species similarity on trait evolution. Here, we consider a model where the evolutionary dynamics of traits involved in interspecific interactions are influenced by species similarity in trait values and where we can specify which lineages are in sympatry. We develop a maximum-likelihood based approach to fit this model to combined phylogenetic and phenotypic data. Using simulations, we demonstrate that the approach accurately estimates the simulated parameter values across a broad range of parameter space. Additionally, we develop tools for specifying the biogeographic context in which trait evolution occurs. In order to compare models, we also apply these biogeographic methods to specify which lineages interact sympatrically for two diversity-dependent models. Finally, we fit these various models to morphological data from a classical adaptive radiation (Greater Antillean Anolis lizards). We show that models that account for competition and geography perform better than other models. The matching competition model is an important new tool for studying the influence of interspecific interactions, in particular competition, on phenotypic evolution. More generally, it constitutes a step toward a better integration of interspecific interactions in many ecological and evolutionary processes.

Evolutionary Biology

Social selection parapatry in an Afrotropical sunbird

The extent of range overlap of incipient and recent species depends on the type and magnitude of phenotypic divergence that separates them. Trait divergence by social selection likely initiates many speciation events, but may yield niche-conserved lineages predisposed to limit each others ranges via ecological competition. Here we examine this neglected aspect of social selection speciation theory in relation to the discovery of a non-ecotonal species border between sunbirds. We find that Nectarinia moreaui and N. fuelleborni meet in a ~6 km wide contact zone, as estimated by molecular cline analysis. These species exploit similar bioclimatic niches, but sing highly divergent learned songs, consistent with divergence by social selection. Cline analyses suggest that within-species stabilizing social selection on song-learning predispositions maintains species differences in song despite both hybridization and cultural transmission in the contact zone. We conclude that ecological competition between moreaui and fuelleborni contributes to the stabilization of the species border, but that ecological competition acts in conjunction with reproductive interference. The evolutionary maintenance of learned song differences in a hybrid zone recommend this study system for future studies on the mechanisms of learned song divergence and its role in speciation.

Evolutionary Biology

Antibiotic Stress Selects against Cooperation in a Pathogenic Bacterium

Ecological antagonisms such as predation, parasitism, competition, and abiotic environmental stress play key roles in shaping population biology, in particular by inducing stress responses and selecting for tolerant or resistant phenotypes. Little is known, however, about their impact on social traits, such as the production of public goods. Evolutionary trade-off theory predicts that adaptation to stresses should lessen investments in costly helping behaviours when cooperation does not increase resistance or tolerance, but support for this prediction is scarce. We employed theory and experiments to investigate how ecological antagonism influences social dynamics and resistance evolution in the pathogenic bacterium Pseudomonas aeruginosa. We subjected two clones of bacterium to four doses of antibiotics and assessed growth and frequencies of public goods producing and non-producing genotypes. Our results show that abiotic stress selects against public goods production. Specifically, we found that non-producers of costly iron chelating molecules (siderophores) most rapidly increased in frequency under intermediate antibiotic pressure. Moreover, the dominance of non-producers in mixed cultures was associated with higher survival and resistance to antibiotics than in either producer or non-producer monocultures. Mathematical modelling explains this counterintuitive result, and shows how these qualitative patterns are predicted to generalise to many other systems. Our results shed light on the complex interactions between social traits and ecological antagonisms, and in particular the consequences for bacterial social evolution and antibiotic resistance.

Microbiology

Cryptic diversity and population structure at small scales: The freshwater snail Ancylus (Planorbidae, Pulmonata) in the Montseny mountain range

Anthropogenic impacts like intensified land use and climate change are severe threats to freshwater biodiversity and effective biodiversity monitoring is therefore one of the most urgent tasks. This is however often hampered by the lack of knowledge regarding the number and ecology of species. Molecular tools have shown many freshwater taxa to comprise morphologically cryptic species, which often occur in sympatry on a small geographic scale. Here, we studied the freshwater snail Ancylus fluviatilis (MUELLER, 1774) species complex in the Iberian Montseny Mountains. We hypothesised 1) that several species of A. fluviatilis sensu lato occur in the Montseny, 2) that different Ancylus species seldom co-occur in syntopy due to different ecological demands or interspecific competition, and 3) that species show a pattern of strong population structure within streams or catchments due to ecological preferences or local adaptation. We barcoded 180 specimens from 36 sites in the Montseny for the cytochrome c oxidase subunit I (COI) barcoding gene and molecularly identified two Ancylus species. These species seldom occurred in syntopy and a species distribution modelling approach showed differing bioclimatic preferences of the species. One species mainly occurs in cooler, higher altitude streams while the second species occurs in lower-altitude areas with higher temperatures. Tests of population structure showed that both species possibly do not disperse well in the study area and that populations within species are likely adapted to certain bioclimatic conditions in different regions of the Montseny. Our results highlight the need to incorporate molecular techniques into routine monitoring programmes.

Molecular Biology