The reliability of estimates of migration in the peppered moth Biston betularia and some implications for selection-migration models

Migration of adult males is one of the important variables involved in the mathematical models of industrial melanism in Biston betularia. Values for this variable are based on data from a capture-recapture performed by Bishop (1972) using both local and bred moths which were at least one night old at release. We carried out an experiment to compare the rate of recapture close to the point of release for moths allowed to fly away immediately after their emergence around dusk and those which were at least one night old at release. Unheld moths were less likely to be recaptured suggesting that males have an initial dispersal phase on their first night which results in a higher rate of emigration than on subsequent nights. Such a phase would have been largely missed in Bishop's experiment. The implications of this type of behaviour pattern for the models of spatial variation based on a selection-migration balance are discussed.     

A decline of melanism in the peppered moth Biston betularia in The Netherlands

Melanic forms of the peppered moth Biston betularia were well established in The Netherlands by the end of the 19th century, indeed the first records of the black carbonaria form in 1867 are only about 20 years later than in England. Analysis of extensive sampling data collected by B. J. Lempke for a period of several years beginning in 1969 shows that carbonaria was at a frequency of about 60 to 70% in most of the country where epiphyte communities on trees were reduced due to the effects of air pollution. The pale typica and the three intermediate insularia forms were each at similar, low frequencies. Only in the extreme north and south-east of The Netherlands where epiphyte floras were richer was carbonaria at a lower frequency of less than 40%. Samples collected from seven localities in 1988 show that carbonaria has dramatically declined to a frequency of less than 10%. In contrast to England, the fully black form is being replaced not only by typica but also by the darkest of the insularia phenotypes. The decline in melanism coincides with a period of decreasing levels of sulphur dioxide and of increasing species diversity of lichens on trees.     

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.     

Behavioural studies on the peppered moth Biston betularia and a discussion of the role of pollution and lichens in industrial melanism

We examined the resting behaviour during pairing and egg-laying of bred female Biston betularia (L.) after they were set free at wooded sites in Somerset and Cardiff. Moths were not released immediately on emergence but were held for three nights to overcome an initial period of flight activity. Females (N= 257) were placed on trunks, and on large and small branches of trees, especially oaks, over periods of several weeks in 1984, 1985 and 1986. Releases were not made within 50 m of any known survivor. One hundred and thirty-one pairings were observed. We also examined oviposition behaviour in a cage experiment using different types of branches. Our observations support Mikkola's earlier conclusion from cage experiments with male moths that the species rests predominantly on branches and shows an appropriately specialized resting attitude, demonstrated here in a series of photographs. Many moths will rest underneath, or on the side of, narrow branches in the canopy. Once released females had settled and paired they only moved quite short distances, but positioning and egg-laying were profoundly influenced by the presence of foliose lichens. Females of all phenotypes show a strong preference to oviposit beneath a thallus of foliose lichens. Their tendency to rest against or close to the lichen during the day could markedly influence their crypsis. In the absence of foliose lichens, eggs are laid in cracks in the bark. The survivorship of released females both in different pairing combinations and during their whole reproductive life-span is analysed. These preliminary data suggest that bird predation can give rise to differential mortality of different pairing combinations and of the phenotypes. This technique provides a powerful method of measuring the relative crypsis and differential mortality of phenotypes with living females in natural resting sites. The significance of our observations on resting behaviour is discussed with regard to the role of air pollution and epiphytes in industrial melanism. Particular attention is given to the causal mechanisms underlying recent declines in the frequency of carbonaria melanics in parts of Britain. One important factor is the new growth made by trees since the marked reduction in particulate air pollution in the 1960s. It is emphasized that we need to know much more about the interactions between pollution, epiphytes and resting backgrounds, especially in the canopy, before we can be confident of our understanding of the evolution of industrial melanism.     

Peppered moth revisited: analysis of recent decreases in melanic frequency and predictions for the future

In a recent review article by Mani published in 1990, it was shown that the changes in the frequency of the morph carbonaria can be explained through a migration-selection model and the model is capable of reproducing the decrease in the carbonaria frequency since the enactment of Clean Air Acts. In this paper new data both for carbonaria and for insularia up to 1990 for West Kirby in NW England and for Cambridge are presented. In the earlier review, the data for West Kirby extended to 1987 and for Cambridge to 1985. Data not previously published for the carbonaria and the insularia frequencies at sites in Northwood in Middlesex, Egham in Surrey and Ringwood in Hampshire are also presented. A migration-selection model was used to predict the change in the frequencies of the melanic morphs up to the year 2010. All the data are compared with the predictions of the model. It is encouraging that the data follow the theoretical predictions.     

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.     

Melanic moths and charges in epiphytic vegetation in north-west England and north Wales

In 1973 the appearance and epiphyte cover of tree trunks was examined along a 125 km cline of melanic frequency in the peppered moth in NW England and N Wales. The sampled sites were resurveyed in 1986. There has been a marked increase in the crustose lichen Lecanora conizaeoides at the eastern end of the transect, an increase in lichen diversity in the middle of the transect and a possible decline in large foliose forms at the western end. These changes are discussed in relation to the eastward shift of the cline in melanic frequencies in the moth.     

A bird’s eye view of the peppered moth

Industrial melanism in Biston betularia is one of the best known examples of the role of natural selection in evolution and has received considerable scrutiny for many years. The rise in frequency of the dark form of the moth (carbonaria) and a decrease in the pale form (typica) was the result of differential predation by birds, the melanic form being more cyptic than typica in industrial areas where the tree bark was darkened by air pollution. One important aspect of early work evaluating the relative crypsis of the forms of B. betularia on tree trunks with different lichen flora was the reliance on human observers. Humans, however, do not have the same visual capabilities as birds. Birds have well‐developed ultraviolet (UV) vision, an important component of their colour processing system that affects many aspects of behaviour, including prey detection. We examined the UV characteristics of the two forms of B. betularia and a number of foliose and crustose lichens. In human visible light the speckled form typica appeared cyptic when seen against a background of foliose lichen, whereas the dark form carbonaria was conspicuous. Under UV light the situation was reversed. The foliose lichens absorbed UV and appeared dark as did carbonaria. Typica, however, reflected UV and was conspicuous. Against crustose lichens, typica was less visible than carbonaria in both visible and UV light. These findings are considered in relation to the distribution and recolonization of trees by lichens and the resting behaviour of B. betularia.     

A theoretical analysis of the morph frequency variation in the peppered moth over England and Wales

The migration-selection model for the spatial and temporal variation of morph frequencies of Biston betularia over England and Wales (Cook & Mani, 1980) has been extended to include effects due to non-visual selection. The parameters for non-visual selection were chosen from the recent determination by Mani (1980) and by Creed et al . (1980). The morph frequencies over England and Wales were obtained through computer simulation and the results were compared with data along the Manchester-Yorkshire, Central Wales-East Anglia and South Wales-London transects. Best fits to the data were obtained by using the non-visual selective values of Mani for carbonaria and modified values of Creed et al . for insularia . It is concluded that the observed polymorphism could be well explained through a balance of migration and visual and non-visual selections.     

A theoretical analysis of the morph frequency variation in the peppered moth over England and Wales

The migration-selection model for the spatial and temporal variation of morph frequencies of Biston betularia over England and Wales (Cook & Mani, 1980) has been extended to include effects due to non-visual selection. The parameters for non-visual selection were chosen from the recent determination by Mani (1980) and by Creed et al. (1980). The morph frequencies over England and Wales were obtained through computer simulation and the results were compared with data along the Manchester-Yorkshire, Central Wales-East Anglia and South Wales-London transects. Best fits to the data were obtained by using the non-visual selective values of Mani for carbonaria and modified values of Creed et al. for insularia. It is concluded that the observed polymorphism could be well explained through a balance of migration and visual and non-visual selections.     

Retelling Experiments: H.B.D. Kettlewell's Studies of Industrial Melanism in Peppered Moths

H. B. D. Kettlewell's field experiments on industrial melanism in the peppered moth, Biston betularia, have become the best known demonstration of natural selection in action. I argue that textbook accounts routinely portray this research as an example of controlled experimentation, even though this is historically misleading. I examine how idealized accounts of Kettlewell's research have been used by professional biologists and biology teachers. I also respond to some criticisms of David Rudge to my earlier discussions of this case study, and I question Rudge's claims about the importance of purely observational studies for the eventual acceptance and popularization of Kettlewell's explanation for the evolution of industrial melanism.     

Industrial Melanism in the Peppered Moth, Biston betularia: An Excellent Teaching Example of Darwinian Evolution in Action

The case of industrial melanism in the peppered moth has been used as a teaching example of Darwinian natural selection in action for half a century. However, over the last decade, this case has come under attack from those who oppose Darwinian evolution. Here, the main elements of the case are outlined and the reasons that the peppered moth case became the most cited example of Darwinian evolution in action are described. Four categories of criticism of the case are then evaluated. Criticisms of experimental work in the 1950s that centered on lack of knowledge of the behavior and ecology of the moth, poor experimental procedure, or artificiality in experiments have been addressed in subsequent work. Some criticisms of the work are shown to be the result of lack of understanding of evolutionary genetics and ecological entomology on the part of the critics. Accusations of data fudging and scientific fraud in the case are found to be vacuous. The conclusion from this analysis of criticisms of the case is that industrial melanism in the peppered moth is still one of the clearest and most easily understood examples of Darwinian evolution in action and that it should be taught as such in biology classes.     

Linkage map of the peppered moth,Biston betularia (Lepidoptera,Geometridae): a model of industrial melanism

We have constructed a linkage map for the peppered moth (Biston betularia), the classical ecological genetics model of industrial melanism, aimed both at localizing the network of loci controlling melanism and making inferences about chromosome dynamics. The linkage map, which is based primarily on amplified fragment length polymorphisms (AFLPs) and genes, consists of 31 linkage groups (LGs; consistent with the karyotype). Comparison with the evolutionarily distant Bombyx mori suggests that the gene content of chromosomes is highly conserved. Gene order is conserved on the autosomes, but noticeably less so on the Z chromosome, as confirmed by physical mapping using bacterial artificial chromosome fluorescence in situ hybridization (BAC-FISH). Synteny mapping identified three pairs of B. betularia LGs (11/29, 23/30 and 24/31) as being orthologous to three B. mori chromosomes (11, 23 and 24, respectively). A similar finding in an outgroup moth (Plutella xylostella) indicates that the B. mori karyotype (n=28) is a phylogenetically derived state resulting from three chromosome fusions. As with other Lepidoptera, the B. betularia W chromosome consists largely of repetitive sequence, but exceptionally we found a W homolog of a Z-linked gene (laminin A), possibly resulting from ectopic recombination between the sex chromosomes. The B. betularia linkage map, featuring the network of known melanization genes, serves as a resource for melanism research in Lepidoptera. Moreover, its close resemblance to the ancestral lepidopteran karyotype (n=31) makes it a useful reference point for reconstructing chromosome dynamic events and ancestral genome architectures. Our study highlights the unusual evolutionary stability of lepidopteran autosomes; in contrast, higher rates of intrachromosomal rearrangements support a special role of the Z chromosome in adaptive evolution and speciation.     

Evolution of resting behaviour in polymorphic 'industrial melanic' moth species

The behaviour of individually marked melanic and typical forms of Allophyes oxyacanthae was followed in four successive tests in an apparatus comprising bark of three different reflectances. In samples of wild populations, melanics showed a moderate preference for dark bark, while typicals did not seem to prefer or avoid this substrate. However, in four of the eight families of known parentage tested, both melanics and typicals preferred to rest on dark bark. These four families were the progeny of dark typicals which had shown a preference for dark bark and were the families containing the darkest typicals. In A. oxyacanthae the variation in resting behaviour is not therefore closely associated with the melanic allele but may be linked to a 'dark typical' allele which, together with other loci, produces polygenic darkening of the normal phenotype. The results with A. oxyacanthae are compared with those obtained with Biston betularia and other moth species.     

Localized industrial melanism in the spittlebug Philaenus spumarius (L.) (Homoptera: Aphrophoridae) in Cardiff docks, south Wales

Twenty-four large adult samples of the meadow spittlebug, Philaenus spumarius , were collected from within a 4 times 3 km area containing Cardiff docks, south Wales. All samples are highly polymorphic for colour and pattern and show marked geographic variation in the combined frequency of the eight melanic morphs (from <15% to > 75%). Maximum melanic phenotype frequencies in both sexes coincide with dock wharfs currently containing large accumulations of pulverized coal and these samples are heavily contaminated with small coal particles. However, interpretation of this association is complicated by the existence, until 1976, of a smokeless fuel factory causing local air pollution immediately adjacent to the area of highest melanic frequency. It is suggested that the association of high levels of melanism in P. spumarius with areas heavily contaminated with particulate pollutants is due to direct selective effects of small particles favouring melanies.     

Selection for cuticular melanism reveals immune function and life-history trade-offs in Spodoptera littoralis

Several insect species show an increase in cuticular melanism in response to high densities. In some species, there is evidence that this melanism is correlated with an up-regulation of certain immune system components, particularly phenoloxidase (PO) activity, and with the down-regulation of lysozyme activity, suggesting a trade-off between the two traits. As melanism has a genetic component, we selected both melanic and nonmelanic lines of the phase-polyphenic lepidopteran, Spodoptera littoralis, in order to test for a causative genetic link between melanism, PO activity and lysozyme activity, and to establish if there are any life-history costs associated with the melanic response. We found that, in fact, melanic lines had lower PO activity and higher lysozyme activity than nonmelanic lines, confirming a genetic trade-off between the two immune responses, but also indicating a genetic trade-off between melanism and PO activity. In addition, we found that lines with high PO activity had slower development rates suggesting that investment in PO, rather than in melanism, is costly.     

Coexistence and error propagation in pre-biotic vesicle models: a group selection approach

Compartmentalization of unlinked, competing templates is widely accepted as a necessary step towards the evolution of complex organisms. However, preservation of information by templates confined to isolated vesicles of finite size faces much harder obstacles than by free templates: random drift allied to mutation pressure wipe out any template that does not replicate perfectly, no matter how small the error probability might be. In addition, drift alone hinders the coexistence of distinct templates in a same compartment. Here, we investigate the conditions for group selection to prevail over drift and mutation and hence to guarantee the maintenance and coexistence of distinct templates in a vesicle. Group selection is implemented through a vesicle survival probability that depends on the template composition. By considering the limit case of an infinite number of vesicles, each one carrying a finite number of templates, we were able to derive a set of recursion equations for the frequencies of vesicles with different template compositions. Numerical iteration of these recursions allows the exact characterization of the steady state of the vesicle population-a quasispecies of vesicles-thus revealing the values of the mutation and group selection intensities for which template coexistence is possible. Within the main assumption of the model-a fixed, finite or infinite, number of vesicles-we find no fundamental impediment to the coexistence of an arbitrary number of template types with the same replication rate inside a vesicle, except of course for the vesicle capacity. Group selection in the form of vesicle selection is a must for compartmentalized primordial genetic systems even in the absence of intra-genomic competition of different templates.