Industrial and non-industrial melanism in the peppered moth, Bistort betularia (L.)

Abstract. 1. Estimates of the relative crypsis of the melanic and typical morphs of B.betularia have been made at fifty-two sites in southern England and south Wales and these estimates were compared with melanic frequencies in samples from these sites.
2. The decrease from east to west in the frequency of the melanic form, carbonaria , and increase in the frequency of a second melanic form, insularia , across the southern part of England and Wales is largely independent of changes in smoke concentration, sulphur dioxide concentration or relative crypsis.
3. When all the available information on the distribution of the melanics of B.betularia is considered, sulphur dioxide concentration is correlated with the geographic variation in carbonaria frequency. This, together with evidence of the close relationship between carbonaria crypsis and sulphur dioxide levels, indicates that outside southern England and south Wales, either selective predation, or some direct selective effect of pollutants, is of major importance in determining the variation in carbonaria frequency.
4. Some features of the pattern of the spread of carbonaria in England and Wales during the last century give reasons for expecting non-industrial selective factors to be of greater importance in determining the distribution of melanics of B.betularia in southern England and south Wales than in the rest of Britain.     

On the selective forces acting in the industrial melanism of Biston and Oligia moths (Lepidoptera: Geometridae and Noctuidae)

Melanic and typical morphs of Biston betularius (L.), Oligia latruncula (D. & S.) and 0. strigilis (L.) made choices between vertical trunks and horizontal branches, sprayed with white and black paints, in a transparent plastic cylinder in natural illumination. The moths settled in exposed positions. In neither Biston nor Oligia did the choice for white/black backgrounds differ between the morphs. Biston moths settle on narrow branches (not on twigs) with the body at right angles to the longitudinal axis of the branch. The Oligias showed an asymmetrical light reaction: one eye is kept in shadow so that they settle as a continuation of an irregularity of the surface, often of a lichen.
In nature, Biston betularius probably rests high up in the canopies, on the under surfaces of horizontal branches. The visual selection acting on the morphs is expected to be less intensive than that measured on tree trunks. The mark-release-recapture results of Kettlewell (1955a, 1956) do not show any qualitative change during the self-determination of the moths but the material is too limited for firm conclusions.
Newly-hatched Biston males take off straight from the trunk where they have expanded their wings but the females may climb higher in the tree. A hypothesis is presented to explain the black-and-white coloration of f. carbonaria : the short-winged moths climbing up the trunks might deter bird predation.     

Selective bird predation on the peppered moth: the last experiment of Michael Majerus

Colour variation in the peppered moth Biston betularia was long accepted to be under strong natural selection. Melanics were believed to be fitter than pale morphs because of lower predation at daytime resting sites on dark, sooty bark. Melanics became common during the industrial revolution, but since 1970 there has been a rapid reversal, assumed to have been caused by predators selecting against melanics resting on today's less sooty bark. Recently, these classical explanations of melanism were attacked, and there has been general scepticism about birds as selective agents. Experiments and observations were accordingly carried out by Michael Majerus to address perceived weaknesses of earlier work. Unfortunately, he did not live to publish the results, which are analysed and presented here by the authors. Majerus released 4864 moths in his six-year experiment, the largest ever attempted for any similar study. There was strong differential bird predation against melanic peppered moths. Daily selection against melanics (s ≈ 0.1) was sufficient in magnitude and direction to explain the recent rapid decline of melanism in post-industrial Britain. These data provide the most direct evidence yet to implicate camouflage and bird predation as the overriding explanation for the rise and fall of melanism in moths.     

Theoretical models of melanism in Biston betularia—a review

The various theoretical models that have been constructed to explain the spatial and temporal changes in the frequency of the melanic morph of the moth Biston betularia are reviewed. The assumptions that are made in these models are discussed. It is shown that these models do explain the gross features of the spatial and temporal distribution of melanic variety in England and Wales. The models have been fairly successful in quantitative prediction of the observed decline in the frequencies of the melanic forms by relating selective differentials to the sulphur dioxide levels. Thus these models do yield a reasonable and consistent picture of the gross pattern of changes in melanic frequencies within the limitation of the available data. The models are robust and can thus accommodate changes to the data on the biology and behaviour of the moth. It is shown that neither heterozygote advantage nor non-Darwinian mechanism need to be invoked to understand the observed evolution of melanism in B. betularia.     

A reversible color polyphenism in American peppered moth (Biston betularia cognataria) caterpillars

Insect body color polyphenisms enhance survival by producing crypsis in diverse backgrounds. While color polyphenisms are often indirectly induced by temperature, rearing density, or diet, insects can benefit from immediate crypsis if they evolve polyphenisms directly induced by exposure to the background color, hence immediately deriving protection from predation. Here, we examine such a directly induced color polyphenism in caterpillars of the geometrid peppered moth (Biston betularia). This larval color polyphenism is unrelated to the genetic polymorphism for melanic phenotypes in adult moths. B. betularia caterpillars are generalist feeders and develop body colors that closely match the brown or green twigs of their host plant. We expand on previous studies examining the proximal cues that stimulate color development. Under controlled rearing conditions, we manipulated diets and background reflectance, using both natural and artificial twigs, and show that visual experience has a much stronger effect than does diet in promoting precise color matching. Their induced body color was not a simple response to reflectance or light intensity but instead specifically matched the wavelength of light to which they were exposed. We also show that the potential to change color is retained until the final (sixth) larval instar. Given their broad host range, this directly induced color polyphenism likely provides the caterpillars with strong protection from bird predation.     

Immune defence,a possible nonvisual selective factor behind the industrial melanism of moths (Lepidoptera)

It is generally believed that industrial melanism in Lepidoptera is mainly caused by differential predation by birds. In polluted areas, melanic individuals are favoured by natural selection because they are better camouflaged than pale moths on lichen‐free and sooty tree trunks. In this article, we show that, in the black arches moth (Lymantria monacha), melanic morphs have a stronger encapsulation response than pale morphs against nylon monofilament implants. This indicates that the melanic and pale morphs differ in the strength of their immune defence. The same chemical precursors and their end product, melanin pigment, are involved in the encapsulation response and in the external coloration. Thus, it seems that there may be two possible, not mutually exclusive, explanations for the frequency changes observed in the industrial melanism of moths. The dominant gene causes an increase in the amount of melanin pigment and its precursors. This increase causes two changes: an intensified immune defence as a form of improved encapsulation ability of foreign objects, and the well‐known protective dark coloration (a case of relational pleiotropy). It seems possible that industrial melanism is a by‐product of selection on the strength of immunity. In the field, these pleiotropic aspects are exceedingly difficult to distinguish from each other, and the factors may even be compensatory. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 831–838.     

Decline of melanism in two British moths: spatial, temporal and inter-specific variation

The decline in industrial melanism over the last quarter century constitutes an exceptional case of an evolutionary change, varying in both time and space, and between species. In Biston betularia and Odontoptera bidentata, the change in melanic frequency is closely replicated at two sites 0.5 km apart. Between seven sites 50-100 km apart, there is heterogeneity in both the speed and timing of change. At sites that were heavily industrialized, the change is faster, from an initially higher frequency, and starts later than at sites which are more rural.We propose a method for estimating systematic change during sigmoid declines in melanic frequencies. This fails to show any significant change over time in selective coefficients. It is concluded that the overall pattern of change has been driven largely by events in the most polluted and industrialized parts of the country. Although migration may contribute to the estimated selective values, natural selection is the only credible explanation for the overall decline.     

Resting site selection in the geometrid moth Phigalia pilosaria (Lepidoptera: Geometridae)

When tested in a cylindrical apparatus, males of both typical and melanic morphs of the nocturnal moth Phigalia pilosaria (Schiffermueller) prefer to rest on light backgrounds. This preference is more pronounced when the moths have a rough surface on which to rest than when it is smooth. The two morphs show a distinct tendency to adopt resting positions close to the boundary between the shades presented in the experiments; this is interpreted as being a result of walking moths stopping at or close to the boundary when approaching it from the light side. The surface texture of the background influences the resting attitude of the moths in relation to vertical.     

Background selection by the peppered moth (Biston betularia Linn.): individual differences

The hypothesis that dimorphically coloured, cryptic moths select appropriate rest sites by comparing their body scales to substrate reflectance was tested using typical and melanic morphs of the peppered moth, Biston betularia (L.). Experiments designed to block the individual's inspection of its inherited colour phenotype do not support Kettlewell's contrast/conflict (self-inspection) hypothesis. Instead, tracking of marked moths over successive days revealed individual differences in rest-site selection which were not related to treatments, experience (imprinting), nor closely to a moth's inherited colour pattern. Differences between family broods indicate that some genetic bias in background selection exists. The production of artificially selected lines with consistent but opposing preferences will allow us to investigate the co-evolution of pattern and behaviour.     

Taking the Peppered Moth with a Grain of Salt

H. B. D. Kettlewell's (1955, 1956) classic field experiments on industrial melanism in polluted and unpolluted settings using the peppered moth, Biston betularia, are routinely cited as establishing that the melanic (dark) form of the moth rose in frequency downwind of industrial centers because of the cryptic advantage dark coloration provides against visual predators in soot-darkened environments. This paper critiques three common myths surrounding these investigations: (1) that Kettlewell used a model that identified crypsis as the only selective force responsible for the spread of the melanic gene, (2) that Kettlewell's field experiments alone established that selection for crypsis was the most important factor in the spread of melanic forms, and (3) that Kettlewell's investigations in an unpolluted wood near Dorset constituted a control for his earlier Birmingham studies (contra Hagen 1993, 1996). This analysis further identifies two features that distinguish manipulative experiments in evolutionary biology from experiments in other contexts. First, experiments in evolutionary biology rest on a wealth of information provided by strictly observational ecological studies; in the absence of such information experiments in evolutionary biology make no sense. Second, there is a trade-off between how much control investigators have over the conditions being studied and how informative the results of the experiment will be with regard to natural populations.     

The peppered moth and industrial melanism: evolution of a natural selection case study

From the outset multiple causes have been suggested for changes in melanic gene frequency in the peppered moth Biston betularia and other industrial melanic moths. These have included higher intrinsic fitness of melanic forms and selective predation for camouflage. The possible existence and origin of heterozygote advantage has been debated. From the 1950s, as a result of experimental evidence, selective predation became the favoured explanation and is undoubtedly the major factor driving the frequency change. However, modelling and monitoring of declining melanic frequencies since the 1970s indicate either that migration rates are much higher than existing direct estimates suggested or else, or in addition, non-visual selection has a role. Recent molecular work on genetics has revealed that the melanic (carbonaria) allele had a single origin in Britain, and that the locus is orthologous to a major wing patterning locus in Heliconius butterflies. New methods of analysis should supply further information on the melanic system and on migration that will complete our understanding of this important example of rapid evolution.     

Moths use multimodal sensory information to adopt adaptive resting orientations

Camouflage conceals animals from predators and depends on the interplay between the morphology and behaviour of animals. Behavioural elements of animals, such as the choice of a resting spot or posture, are important for effective camouflage, as well as the animals’ cryptic appearance. To date, the type of sensory input that mediates resting site choice remains poorly understood. Previously, we showed that bark‐like moths perceive and rely on bark structure to seek out cryptic resting positions and body orientations on tree trunks. In the present study, we investigated the sensory organs through which moths perceive the structure of bark when positioning their bodies in adaptive resting orientations. We amputated (or blocked) each one of the hypothetical sensory organs in moths (antennae, forelegs, wings, and eyes) and tested whether they were still able to perceive bark structure properly and adopt adaptive resting orientations. We found that visual information or stimulation is crucial for adaptively orienting their bodies when resting and tactile information from wings may play an additional role. The present study reveals multimodal information use by moths to achieve visual camouflage and highlights the sensory mechanism that is responsible for the adaptive behaviour of cryptic insects. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111 , 900–904.     

The rise and fall of the Carbonaria form of the peppered moth

The evidence for change in frequency of the melanic carbonaria morph in the peppered moth Biston betularia (L.) (Lepidoptera: Geometridae) in England and Wales is reviewed. At mid-20th century a steep cline of melanic phenotype frequency running from the north of Wales to the southern coast of England separated a region of 5% or less to west from 90% or more to northeast. By the 1980s the plateau of 90% frequency had contracted to northern England. The frequency has since continued to drop so that the maximum is now less than 50% and in most places below 10%. There have been similar declines in Europe and North America. Evidence from surveys and from two-point records shows the change to require 5% to 20% selection against the melanic. The melanic is more disadvantageous in regions where its frequency was initially high than in regions where it was low. Experiments to investigate predation by birds show a net advantage to carbonaria morphs in regions where typical frequencies were low at the time of the experiment, and a disadvantage where typical frequencies were high. This would be expected if environment and frequency were associated, and selective predation played a part in generating the association. The cryptic advantage of carbonaria was large in areas of heavy pollution where typical frequencies were 20% or less. The moth usually has a low density but is relatively highly mobile. The ability of present information to explain the patterns has been tested in simulations. They indicate a system under strong selection that has always been in a dynamic state without equilibria.     

Resting orientation enhances prey survival on strongly structured background

Prey can use various camouflage types as defense against predators. One of the most common and important types is background matching, which occurs if an animal matches the background in color, brightness, and pattern. Although background matching has been studied intensively, the effects of the resting orientation of prey on the effectiveness of camouflage through background matching are not well known in natural conditions. Several past experimental studies have been conducted on resting orientation in the lab often using the visual system of humans. Their results revealed that the detection rates of predators hinge on the combination of the resting orientation of artificial moths and their background. Here, we studied whether survival rates of artificial moth-like models depend on their resting orientation in the wild where the visual conditions and detection distances vary. We used a 2 × 2 design of two resting positions of a horizontally and a vertically striped morph on tree bark. Our results show that the survival probability of moths depended mainly on the orientation of stripes relative to the vertical structure of tree bark. Thus, resting orientation relative to background affected survival. After reanalyzing Endler’s (Biol J Linn Soc 22:187–231, 1984) data on resting habitats of 317 species of North American moths, we found that horizontally striped moths occurred frequently on small herbs and tree bark. We suggest that it would be beneficial for striped moths to orient non-randomly on strongly structured background, like furrows of tree bark. We further suggest that background matching was more important than coincident disruptive coloration in determining the survival rates of our artificial moths.     

Melanic frequencies in three species of moths in post industrial Britain

Changing patterns of morph frequency in three moth species ( Biston betularia , Odontoptera bidentata and Apocheima pilosaria ) have been investigated using data from the Rothamsted Insect Survey. All three exhibited industrial melanism during the period of high atmospheric pollution in Britain. Three historical and habitat types are compared, the old industrial north of England, rural Scotland, Wales and South-West England, and a southern English intermediate region of high human population density but generally low industrialization. Between 1974 and 1999 the carbonaria morph of B. betularia declined in frequency in the industrial region and is nearly absent from rural areas. It is the form which most closely tracks atmospheric change. It is shown that the insularia forms of B. betularia and the melanic morphs in the other two species have decreased in the industrial region, commencing later than carbonaria , but have maintained their presence and possibly reached equilibrium elsewhere. They may be non-industrial polymorphisms. B. betularia is rarer than the other species and all three species are at lower densities in industrial than in non-industrial regions. © 2002 The Linnean Society of London , Biological Journal of the Linnean Society , 2002, 75 , 475–482.     

Experiments on resting site selection by the typical and melanic forms of the moth, Allophyes oxyacanthae (Caradrinidae)

Six families of the cryptic moth, Allophyes oxyucanthae , four of which contained both the typical and melanic forms of this polymorphic species, were tested for evidence of resting site selection in a large box lined with oak bark of three reflectances. Moths from different families were tested separately. Typicals from all four families tended to show a preference for the background which most closely matched their reflectance and on which they were most cryptic. Melanic moths from different families showed different resting site selection behaviour; those from two families preferring dark bark, on which melanics were most cryptic, while those from other families did not. The significance of these results in relation to previous suggestions about the control of resting site selection is discussed     

The costs of colour: plasticity of melanin pigmentation in an outbreaking polymorphic forest moth

Outside the context of industrial melanism, little is known about the physiological and ecological importance of genetic melanic polymorphisms in moths. Melanin pigments are synthesized from amino acid precursors and should therefore be costly to produce in nitrogen‐limited insects. A genetic melanic polymorphism is present in adult Malacosoma disstria Hübner (Lepidoptera: Lasiocampidae), a widespread forest moth with outbreaking population dynamics. We test the hypotheses that melanin‐based colouration is physiologically costly in M. disstria, that expression of melanin‐based colouration is a plastic trait which varies with population density and nutrition, and that the genetically based melanic phenotype is disadvantaged under nutritionally poor conditions. Two experiments were used to test these hypotheses. A field study compared pigmentation and phenotypic frequencies in moths collected from high‐ and low‐density populations. A laboratory experiment investigated the effects of larval nitrogen availability on adult pigmentation and phenotypic frequencies. High population density and nitrogen limitation reduced pigmentation and size of all moths, but phenotypic frequencies were not affected in either experiment. The effects of diet on both pigmentation and size were stronger for melanic moths than for typical moths. Our results show that adult melanism in M. disstria is physiologically costly, that colour expression is plastic despite its genetic component, and that the melanic phenotype may be disadvantaged under poor conditions but favoured under good conditions. We suggest that temporal variation in selection and trait plasticity help maintain polymorphism stability.     

Industrial melanism and peppered moths (Biston betularia (L.))

The spread of melanic forms of the peppered moth ( Biston betularia (L.)) over polluted areas of Britain from the mid-nineteenth century onwards, has become widely known and quoted as a classical example of microevolutionary change. Probably the most important factor in the spread (and subsequent decline, following the Clean Air Act) of the melanics has been bird predation on less cryptic individuals, but a range of other factors may also affect the maintenance of allele frequencies at any one place (site selection, dispersion, heterosis, frequency dependent selection, larval hardiness, etc). The development of the "Peppered Moth Story" is described, and suggestions made about needed research.     

Differential winter mortality and seasonal selection in the polymorphic ladybird Adalia bipunctata (L) in the Netherlands

Seasonal selection acting on the melanic polymorphism in the two-spot ladybird Adalia bipunctata was investigated in The Netherlands. An increase in melanic frequency over the spring-summer reproductive period was quantified. The selective advantage gained by melanics averaged 9%, but significant heterogeneity occurred between populations. Adult hibernation behaviour is described. The beetles when outdoors show a highly clumped distribution both between and within trees. The distribution of the morph classes between aggregations is random. Survivorship in a hibernating cohort (initial n= 1898) on a grid of 70 lime trees near Utrecht was monitored by making three counts over the winter of 1981–1982. Intense selection favouring each melanic morph occurred during December and January. The relative fitness of non-melanics was 0.55 (melanics =1). The discovery of dead beetles in late January (about 5% of total losses) and the absence of spatially density-dependent mortality were consistent with a climatic stress rather than selective predation. The period of selection was associated with very cold temperatures averaging up to 4°C below normal and an overall mortality of nearly 75%. There was no change in morph frequency, near normal temperatures and a lower mortality from February to early April. Examination of groups of nearby trees in late January strongly suggested that similar differential mortality had occurred except on some willows. This difference was probably due to the more protected hibernation sites available on these trees. Samples of hibernating cohorts at three other sites showed no evidence of differential mortality. Laboratory experiments with hibernating beetles found no difference in survivorship or rate of weight loss between starved non-melanics and melanics in temperature regimes with and without periods of adult activity. It is concluded that the intense winter selection on the study limes is probably exceptional. Examination of changes in morph frequency through the annual cycle suggests that at some sites the selection favouring melanics during reproduction is counterbalanced by selection against melanics in late summer or early autumn. The results are discussed in relation to mathematical models of cyclical selection and to other field studies including that of Timoféeff-Ressovsky (1940), who found large decreases in melanic frequency during hibernation in Berlin.     

Changing views on melanic moths

The rapid rise in frequency of melanic morphs in several moth species, especially the peppered moth Biston betukria , in industrial regions during the 19th century, and the subsequent rapid decline, indicate the action of strong selection. There has recently been a tendency to criticise and question all aspects of research on industrial melanism, including the experiments which suggest that selective predation plays an important part in the changes. These experiments are reexamined, together with evidence for changes in appearance of tree surfaces and for relation of initial melanic frequency to subsequent rate of decline. It is suggested that intense pollution may have been required to drive the carbonaria morph to a high frequency, with frequency patterns over a mosaic environment smoothed by migration. Improvements in these extreme locations then triggered the decline, with litde indication of the environmental changes in areas of moderate pollution. Reasons for criticism of past work are discussed. Industrial melanism continues to provide an exceptional opportunity to analyse a pattern of selection and change in gene frequency.