Evolutionary constraints of warning signals: A genetic trade-off between the efficacy of larval and adult warning coloration can maintain variation in signal expression.

To predict evolutionary responses of warning signals under selection, we need to determine the inheritance pattern of the signals, and how they are genetically correlated with other traits contributing to fitness. Furthermore, protective coloration often undergoes remarkable changes within an individual's lifecycle, requiring us to quantify the genetic constraints of adaptive coloration across all the relevant life stages. Based on a 12 generation pedigree with > 11,000 individuals of the wood tiger moth (Arctia plantaginis), we show that high primary defense as a larva (large warning signal) results in weaker defenses as adult (less efficient warning color), due to the negative genetic correlation between the efficacy of larval and adult warning coloration. However, production of effective warning coloration as a larva did not incur any life-history costs and was positively genetically correlated with reproductive output. These results provide novel insights into the evolutionary constraints on protective coloration in animals, and explain the maintenance of variation in the signal expression despite the strong directional selection by predators. By analyzing the genetic and environmental effects on warning signal and life-history traits in all relevant life stages, we can accurately determine the mechanisms shaping the evolutionary responses of phenotypic traits under different selection environments.

Different food sources elicit fast changes to bacterial virulence.

Environmentally transmitted, opportunistic bacterial pathogens have a life cycle that alternates between hosts and environmental reservoirs. Resources are often scarce and fluctuating in the outside-host environment, whereas overcoming the host immune system could allow pathogens to establish a new, resource abundant and stable niche within the host. We tested if short-term exposure to different outside-host resource types and concentrations affect Serratia marcescens-(bacterium)'s virulence in Galleria mellonella (moth). As expected, virulence was mostly dictated by the bacterial dose, but we also found a clear increase in virulence when the bacterium had inhabited a low (versus high) resource concentration, or animal-based (versus plant-based) resources for 48 h prior to injection. The results suggest that temporal changes in pathogen's resource environment can induce very rapid changes in virulence and affect infection severity. Such changes could also play an important role in shifts from environmental lifestyle to pathogenicity or switches in host range and have implications for the management of opportunistic pathogens and disease outbreaks.

Costs and benefits of plant allelochemicals in herbivore diet in a multi enemy world.

Sequestration of plant defensive chemicals by herbivorous insects is a way of defending themselves against their natural enemies. Such herbivores have repeatedly evolved bright colours to advertise their unpalatability to predators, i.e. they are aposematic. This often comes with a cost. In this study, we examined the costs and benefits of sequestration of iridoid glycosides (IGs) by the generalist aposematic herbivore, the wood tiger moth, Parasemia plantaginis. We also asked whether the defence against one enemy (a predator) is also effective against another (a parasitoid). We found that the larvae excrete most of the IGs and only small amounts are found in the larvae. Nevertheless, the amounts present in the larvae are sufficient to deter ant predators and also play a role in defence against parasitoids. However, excreting and handling these defensive plant compounds is costly, leading to longer development time and lower pupal mass. Interestingly, the warning signal efficiency and the amount of IGs in the larvae of P. plantaginis are negatively correlated; larvae with less efficient warning signals contain higher levels of chemical defence compounds. Our results may imply that there is a trade-off between production and maintenance of coloration and chemical defence. Although feeding on a diet containing IGs can have life-history costs, it offers multiple benefits in the defence against predators and parasitoids.

Low parasitism rates in parthenogenetic bagworm moths do not support the parasitoid hypothesis for sex.

The parasite hypothesis for sex is one of the many theories that have been suggested to solve the mystery of the widespread occurrence of sex despite its high short-term costs. It suggests that sexual lineages have an evolutionary advantage over parthenogens because they can frequently generate new genotypes that are temporarily less prone to coevolving parasites. In this study, we looked for further supporting evidence for the parasite hypothesis of sex in an attempt to understand the coexistence of sexual and parthenogenetic bagworm moths (Naryciinae). The bagworm moths and their parasitoids form one of the few natural host-parasite systems where sexual and parthenogenetic hosts are apparently not separated by ecological or geographical barriers. Furthermore, in support of the parasite hypothesis for sex, parthenogenetic presence is negatively correlated with parasitism rate. We specifically tested, by identifying the reproductive mode of the parasitized individuals, whether parasitoids preferentially attack the parthenogens in sites with both sexual and parthenogenetic forms, as predicted by the parasite hypothesis. We collected hosts from sites with different frequencies of parthenogenetic and sexual moths. A DNA barcoding approach was used to determine the reproductive mode of the parasitized hosts. Furthermore, we investigated whether differences in host and parasitoid phenology could provide an alternative explanation for the variation in parasitism rates between parthenogens and sexuals. Our results contradict the prediction of the parasite hypothesis because parthenogenetic bagworm moths were less parasitized than sexuals in sympatric sites. Our findings can be explained by differences in phenology between the parthenogenetic and sexual moths rather than genetic incompatibility between parthenogenetic hosts and parasitoids. The stable coexistence of sexual and parthenogenetic Naryciinae despite the many apparent costs of sex in this system remains a mystery. Our work adds to the list of studies were the assumptions of the parasite hypothesis for sex are not all met.

Investigating Müllerian mimicry: predator learning and variation in prey defences.

Inexperienced predators are assumed to select for similarity of warning signals in aposematic species (Müllerian mimicry) when learning to avoid them. Recent theoretical work predicts that if co-mimic species have unequal defences, predators attack them according to their average unpalatability and mimicry may not be beneficial for the better defended co-mimic. In this study, we tested in a laboratory environment whether a uniform warning signal is superior to a variable one in promoting predator learning, and simultaneously whether co-mimics are preyed upon according to their average unpalatability. There was an interaction of signal variation and unpalatability but inexperienced birds did not select for signal similarity in artificial prey; when the prey was moderately defended a variable signal was even learnt faster than a uniform one. Due to slow avoidance learning, moderately defended prey had higher mortality than highly defended prey (although this was not straightforward), but mixing high and moderate unpalatability did not increase predation compared with high unpalatability. This does not support the view that predators are sensitive to varying unpalatability. The results suggest that inexperienced predators may neither strongly select for accurate Müllerian mimicry nor affect the benefits of mimicry when the co-mimics are unequally defended.

Negatively condition dependent predation cost of a positively condition dependent sexual signalling.

Predation is considered as an important factor constraining the expression of sexual signals. Nevertheless, direct quantitative evidence for predation provoking significant viability costs on individuals signalling at high rates is scarce. Moreover, it is unclear whether high rate signallers are able to balance presumably increased predation costs. We examined whether a condition dependent audible sexual signal, drumming, makes Hygrolycosa rubrofasciata male spiders more prone to predation by pied flycatchers (Ficedula hypoleuca), and whether sexual signalling rate is related to escaping ability once attacked. When birds were given a choice between two spider males manipulated to drum either one or three bouts per minute using playbacks, naïve birds attacked the males randomly regardless of the drumming rate. However, experienced birds chose significantly more often the males with high signalling rate. When spiders were allowed to escape, males with high sexual signalling rate tended to be better at escaping attacks than males with low sexual signalling rate. This study provides evidence that high signalling rate increases the risk of predation, but simultaneously males with high mobility, which correlates positively with signalling rate seemed to be better at compensating this cost.

Multiple benefits of gregariousness cover detectability costs in aposematic aggregations.

Understanding the early evolution of aposematic (warning) coloration has been a challenge for scientists, as a new conspicuous morph in a population of cryptic insects would have a high predation risk and would probably die out before local predators learnt to avoid it. Fisher presented the idea of aggregation benefit through the survival of related individuals; however, his theory has been strongly debated as the mechanisms that favour grouping have never been explored experimentally with the incorporation of detectability costs. Here we create a comprehensive 'novel world' experiment with the great tit (Parus major) as a predator to explore simultaneously the predation-related benefits and costs for aposematic aggregated prey, manipulating both group size and signal strength. Our results show that grouping would have been highly beneficial for the first aposematic prey individuals surrounded by naive predators, because (1) detectability risk increased only asymptotically with group size; (2) additional detectability costs due to conspicuous signals were marginal in groups; (3) even naive predators deserted the group after detecting unpalatability (dilution effect); and (4) avoidance learning of signal was faster in groups. None of these mechanisms require kin selection.

Selection for cryptic coloration in a visually heterogeneous habitat.

We studied selection by predators for cryptic prey coloration in a visually heterogeneous habitat that consists of two microhabitats. It has been suggested that the probability of escaping detection in such habitats might be optimized by maximizing crypsis in one of the microhabitats. However, a recent model indicates that a coloration that compromises the requirements of different microhabitats might sometimes be the optimal solution. To experimentally study these hypotheses, we allowed great tits (Parus major L.) to search for artificial prey items in two different microhabitats (background boards): small patterned and large patterned. On each board there was one prey item that was either small-patterned, large-patterned or medium-patterned and thus compromised. Search time was used as the measure of crypsis and was on average longer on the large-patterned than on the small-patterned background. On the small-patterned background, the small-patterned prey was more cryptic than the compromised prey, which was in turn more cryptic than the large-patterned prey. On the large-patterned background, the small-patterned prey was least cryptic, but the compromised prey did not differ significantly from the large-patterned prey. The compromised coloration had lower predation risk than the matching colorations. This indicates that in some conditions a compromised coloration might be the best strategy for the prey and has important implications for the study of animal coloration.

Strong antiapostatic selection against novel rare aposematic prey.

The evolution of aposematism, a phenomenon where prey species conspicuously advertise their unprofitability to predators, is puzzling. How did conspicuousness evolve, if it simultaneously increased the likelihood of an inexperienced predator to detect the prey and presumably kill it? Antiapostatic selection, where rare prey is predated relatively more often, is considered as another major difficulty for aposematism to evolve. However, the risk of being conspicuous in low frequencies has not been experimentally tested. We designed an experiment to test how frequency (4%, 12%, 32%) of conspicuous aposematic prey and its dispersion type (solitary vs. aggregated) affect an initial predation risk of the prey and in avoidance learning of predators. Wild great tits (Parus major) were predators on artificial prey in a "novel world." The relative mortality of aposematic prey was antiapostatic, thus the frequency-dependent predation was most severe at low frequencies. In all frequencies, aggregated aposematic prey survived better than solitary prey. Surprisingly, learning was not determined by a fixed number of unpalatable prey eaten, but at low frequencies fewer aposematic individuals eaten generated predators' avoidance learning. However, per-capita risk for the prey remained highest at low frequencies. Our results underscore the problems of initial evolution of rare conspicuous morphs. Aggregated prey suffered less from predation, indicating selective advantage of aggregation over solitary living for a conspicuous individual.

Predator experience on cryptic prey affects the survival of conspicuous aposematic prey.

Initially, aposematism, which is an unprofitable trait, e.g. noxiousness conspicuously advertised to predators, appears to be a paradox since conspicuousness should increase predation by naive predators. However, reluctance of predators for eating novel prey (e.g. neophobia) might balance the initial predation caused by inexperienced predators. We tested the novelty effects on initial predation and avoidance learning in two separate conspicuousness levels of aposematic prey by using a 'novel world' method. Half of the wild great tits (Parus major) were trained to eat cryptic prey prior to the introduction of an aposematic prey, which potentially creates a bias against the aposematic morph. Both prey types were equally novel for control birds and they should not have shown any biased reluctance for eating an aposematic prey. Knowledge of cryptic prey reduced the expected initial mortality of the conspicuous morph to a random level whereas control birds initially ate the conspicuous morph according to the visibility risk. Birds learned to avoid conspicuous prey in both treatments but knowledge of cryptic prey did not increase the rate of avoidance learning. Predators' knowledge of cryptic prey did not reduce the predation of the less conspicuous aposematic prey and additionally predators did not learn to avoid the less conspicuous prey. These results indicate that predator psychology, which was shown as reluctance for attacking novel conspicuous prey, might have been important in the evolution of aposematism.

Acoustic signalling in a wolf spider: can signal characteristics predict male quality?

While there has been considerable interest in female choice for male sexual signals, there have been few studies of the underlying information that different aspects of the signal calls convey. Such studies, however, are essential to understand the significance of signals as honest handicaps, arbitrary Fisherian traits and/or in species recognition. We studied the somewhat exceptional system of audible drumming in the wolf spider Hygrolycosa rubrofasciata. We estimated the repeatabilities of signal components, the levels of between-male variance, the symmetry of the signal, the correlations between different aspects of drumming and their correlations with body weight. While in other taxa the frequency of audible signals may convey honest information of male size, in this species signal frequency was not related to male size and had a low repeatability. The pulse rate within each drum was highly repeatable but had a relatively small between-male coefficient of variation. In previous studies on this species, these traits were not important for male mating success. Among the traits directionally preferred by females, signal volume had considerable repeatability. Signal length was repeatable with high variability between males. In one population, signal length and volume were positively correlated with the rate at which males produced the drumming signals, a trait essential for male mating success. Thus, while signal length may reliably indicate male quality, other signal characteristics such as peak frequency and symmetry were not repeatable or were static and not related to any other male traits. Copyright 2000 The Association for the Study of Animal Behaviour.

Maternal effort and male quality in the bank vole, Clethrionomys glareolus.

Parental investment in reproduction is adjusted according to potential benefits in terms of offspring survival and/or mating success. If male quality affects the reproductive success of a female, then females mating with high-quality males should invest more in reproduction. Although the subject has been of general interest, further experimental verification of the hypothesis is needed. We studied whether female bank voles (Clethrionomys glareolus) adjusted their maternal effort according to male quality, measured as mating success. To enable the measurement of maternal effort during nursing separately from male genetic effects the litters were cross-fostered. Further, the genetic background of male quality was examined. Male quality did not correlate with litter size or offspring size at birth. Offspring growth was positively related to food consumption and milk production of mothers. However, these direct measurements of maternal effort were independent of male quality. Male mating success appeared to be significantly heritable indicating that there are genetic benefits. Still, females did not adjust maternal effort according to the genetic quality of their offspring. We suggest that female bank voles gain significant genetic benefits from mating with high-quality males whereas they cannot improve their reproductive success by increasing maternal effort.

Ectoparasites, nest site choice and breeding success in the pied flycatcher.

It has recently been suggested that nest box studies might bias the measurement of behavioural and life-history traits, because the removal of old nests may reduce the load of ectoparasites. This experimental artefact may have notable effects on nest site choice and breeding success in cavity-breeding birds. We tested (i) if pied flycatchers Ficedula hypoleuca prefer clean nest boxes and (ii) if old nest material affects the number of parasites and the breeding success of pied flycatchers. In the first experiment we offered birds one cleaned nest box and one nest box with old nest material from the previous year. The two nest boxes were placed in very similar sites near each other. In this experiment all pied flycatchers clearly preferred "dirty" nest boxes. In the second part of the study we distributed clean nest boxes and dirty ones on the same study area. After breeding was over we counted the number of fleas Ceratophyllus gallinae in the nest material. This flea species was the most abundant and probably the most hazardous parasite in the nests. Surprisingly, we found that there were significantly more fleas in the nest boxes with nests of the current year only than in the boxes with nests of both current and previous year. This might explain the preference for the dirty boxes. However, our results do not indicate that the number of fleas affects breeding success in the pied flycatcher.