Animal Behaviour 82 (2011) 1143e1149

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Facultative social parasites mark host nests with branched hydrocarbons
M. Cristina Lorenzi a, *, Rita Cervo b,1, Anne-Geneviève Bagnères c, 2

Dipartimento di Biologia Animale e dell’Uomo, Università di Torino Dipartimento di Biologia Evoluzionistica Leo Pardi, Università di Firenze c I.R.B.I., UMR CNRS 6035, Université de Tours

a r t i c l e i n f o
Article history: Received 26 May 2011 Initial acceptance 23 June 2011 Final acceptance 8 August 2011 Available online 13 September 2011 MS. number: 11-00433R Keywords: brood parasitism cuticular hydrocarbon nest usurpation Polistes biglumis scent marking social wasp

The chemical integration strategies of facultative social parasites of social insects have not received the scientific attention they deserve, even though there is considerable research being done on the strategies of obligate social parasites. We simulated intraspecific nest usurpations in the social paper wasp, Polistes biglumis, by dividing each nest into two parts and putting one half in the care of the original foundress and the other half in the care of a usurper. After 8 days, we removed and killed foundresses and usurpers, and later tested the responses of naïve, sister-offspring to them. In each half-colony, the offspring were more tolerant to the female that was last on the nest, regardless of whether she was the foundress or a usurper. This suggested that usurpers had the chemical means to be tolerated by the host offspring. Comparisons between the epicuticular hydrocarbon profiles of foundresses and usurpers showed that usurpers were neither chemically insignificant nor transparent, nor were they mimetic, as obligate parasites often are. Instead, usurpers had chemical profiles richer in methyl-branched hydrocarbons than those of the foundresses. Analyses of the hydrocarbon profiles of nest paper revealed that usurpers supplemented host nests with their own hydrocarbons, a sort of nest marking. As a result, the chemical profiles of the host nests became qualitatively more similar to those of the usurpers. These chemical strategies illustrate that branched hydrocarbons play a role as semiochemicals and that facultative parasites may not all be on the main pathway to obligate parasitism. Ó 2011 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

Parasites exploit any resources that free-living organisms offer: from molecular engines to whole cells, from whole organisms to the structures they make. In their struggle to exploit hosts, parasites provide some of the best evidence of adaptation by natural selection. Brood parasites are species that do not target organisms but rather the resources that free-living organisms build or produce: their nests, their parental care and/or their social structures (Wilson 1971; Rothstein 1990). Brood parasites exhibit traits that give them an advantage in fooling and exploiting their hosts and these traits are the results of reciprocal hosteparasite interactions across evolutionary time (Brooke & Davies 1988). The recognition systems of birds and social insects protect nests from brood parasites. For example, the hosts of cuckoos discriminate between their own eggs and those of cuckoos visually, and so cuckoos lay visually mimetic eggs (Rothstein 1990; Rothstein & Robinson 1998). Furthermore, social insects distinguish nestmates
* Correspondence: M. C. Lorenzi, Dipartimento di Biologia Animale e dell’Uomo, Università di Torino, Via Accademia Albertina 13, 10123 Torino, Italy. E-mail address: (M. Cristina Lorenzi). 1 R. Cervo is at the Dipartimento di Biologia Evoluzionistica Leo Pardi, Università di Firenze, Via Romana 17, 50125 Firenze, Italy. 2 A.-G. Bagnères is at the I.R.B.I., UMR CNRS 6035, Université de Tours, Faculté des Sciences, Parc Grandmont, 37200 Tours, France.

from non-nestmates by means of chemicals, and social insect parasites (social parasites) trick their hosts about their own chemical identity (Bagnères & Lorenzi 2010). Doing so, social parasites enter host colonies, exploit host nests and use the host workforce for their own reproduction. The information about chemical identity in social insects is conveyed by epicuticular hydrocarbon blends (Howard & Blomquist 2005; Blomquist & Bagnères 2010). Generally, individuals from different species have epicuticular hydrocarbon blends that differ in composition (Bagnères & Wicker-Thomas 2010). Within species, individuals from different colonies have hydrocarbon blends that differ in the relative proportions of their compounds (BonavitaCougourdan et al. 1987; Bruschini et al. 2010; Van Zweden & d’Ettorre 2010). Within colonies, nestmates recognize each other because their chemical profiles are very similar. Social parasites escape detection by hosts in at least three ways: they mimic the chemical profiles of their hosts (chemical mimicry, sensu Dettner & Liepert 1994); they have chemical profiles that are poor in hydrocarbons (chemical insignificance, Lenoir et al. 2001); and/or they lack some of the hydrocarbons of their hosts (chemical transparency, Martin et al. 2008a). One illustrative example of chemical mimicry is that of the slavemaker Polyergus queens, which take on chemical profiles that

0003-3472/$38.00 Ó 2011 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.anbehav.2011.08.011

Johnson et al. We noted the occurrence of the following behaviours: biting. We used them later in behavioural tests and chemical analyses (see below). Buschinger 2009). Each half comb was separately fastened to the wall of a plastic box (18 Â 12 cm and 11 cm high) and either its original foundress (foundress half-colony) or a usurper (usurper half-colony) was put into the box. P ¼ 0. General Procedure We split nests collected in the field into two parts: one half was reared by the original foundress (foundress half-colonies) and the other half was ‘put up for usurpation’ to an alien female (usurper. the phase when the foundresses are the only adults in the colonies. facultative social parasites (hereafter usurpers) invade host colonies during the founding phase. Lorenzi et al. the offspring never met their foundresses or usurpers during adulthood and were immature in the 8-day period that foundresses and usurpers spent on the half-colonies. Therefore the offspring were naïve to foundresses and usurpers when we performed the behavioural tests. Lorenzi & Bagnères 2002. Therefore we expected that usurpers would not be related to the colony they usurped. 1997. Unlike most obligate parasites. attempting to sting and grasping. The half-colonies were kept under a 12:12 h light:dark photoperiod and supplied with honey. Cervo 2006). 2001. 2001). Wilcoxon pairwise test: Z ¼ À0. In the behavioural tests. all three species of the obligate social parasite Polistes wasps perfectly mimic their congeneric host chemical profiles (Bagnères et al. D’Ettorre & Errard 1998. the usurper of their own half-colony or of their sisters’ half-colony) by performing bioassays using the dead foundresses and usurpers.1144 M. facultative.962. At room temperature. We collected 24 colonies in Chalpe (Italian Western Alps: 44 560 N. This function was probably coopted by a semeiotic function in a series of evolutionary steps that are mostly unknown (Le Conte & Hefetz 2008). usurpers attack. Behavioural Tests We tested whether the offspring discriminated between their foundress (i. usurper half-colonies: 16e28 cells. In Polistes. water and Tenebrio molitor larvae ad libitum. social parasites may contribute to identifying the steps towards the evolution of the chemical strategies that obligate parasites use to overcome host detection. Z ¼ À0.33 Æ 0. Intraspecific facultative parasitism is common among social insects (Hölldobler & Wilson 1990. In the social wasp Polistes biglumis Linnaeus. the singly founded colonies are often the targets of conspecific usurpers (Lorenzi & Cervo 1995). up to 18% of the colonies are usurped by conspecific usurpers. The first offspring emerging from the foundress halfcolonies were sisters to those emerging from the usurper halfcolonies and both were daughters of their original foundresses (usurpers usually keep the first offspring of the foundress when they usurp host colonies. Usurpers and ‘host’ colonies came from two populations separated by mountain barriers. / Animal Behaviour 82 (2011) 1143e1149 match those of their Formica hosts (Habersetzer & BonavitaCougourdan 1993. 2000. possibly after they have lost their own nests (Lorenzi & Cervo 1995). Wilcoxon pairwise test: Z ¼ À1. Within a colony. Lorenzi et al. its usurper came from the other.420. we waited for their emergence. The offspring tested did not differ significantly in age between half-colonies (on average: 3. chase away or kill adult hosts. Turillazzi et al. biglumis colonies in Montgenèvre (French Hautes Alpes: 44 550 N. usurper half-colonies). 1987. we randomly chose 12 colonies from each collection site. These chemical strategies can only be explained as the result of hosteparasite coevolution (Lorenzi 2006. we introduced the foundress and the usurper separately into each pair of half-colonies in random order and at least 1 h apart.71 days old in foundress half-colonies. If the colony came from one population. usurpers dominate them and force them into rearing their own brood (Cervo & Dani 1996. Cervo 2006. In the laboratory. We investigated whether usurpers are chemically insignificant. we know very little about the evolutionary steps that led social parasites to evolve chemical insignificance and chemical mimicry. basic function of insect hydrocarbons was probably physical: the limitation of desiccation. hydrocarbon profiles differ between colonies (Lorenzi et al.44 adult female offspring per usurper half-colony. Cervo & Lorenzi 1996). Then usurpers take over host colonies. Because foundress and usurper half-colonies had immature offspring. 6 430 E).826.38 Æ 2. When the host brood emerge. Cristina Lorenzi et al. mimetic or transparent by simulating intraspecific nest usurpation in the laboratory. The observer did not know whether the dead female was the foundress or the usurper in the test half-colony. foundress half-colonies: 5e8 brood.72 days old in usurper halfcolonies. the targeted colonies are solitary foundations and hence a single foundress is the only adult defending her colony (Cervo & Dani 1996. usurper half-colonies: 2e8 brood. Bioassays using dead insects eliminate the confounding effects of behavioural or chemical actions by the insects introduced and are routinely used in recognition experiments in social insects (e. Lorenzi et al.g. the hydrocarbon profiles of adult wasps are similar to each other and to the profiles that cover the paper nest surfaces (Lorenzi et al. Although limited to one species. In this respect.18 Æ 2. 6 490 E) in mid-July 1997. In contrast. we also hope this study broadens our knowledge of female scent marking in animal conflicts in species other than paper wasps. The Polyergus parasite species and two of the Polistes species are also chemically insignificant when they invade host colonies (Lenoir et al. METHODS Model Species In the solitarily founding species P. Similarly. Likewise. we also collected 24 P.18 adult female offspring per foundress half-colony and by 1. 6. The colonies were towards the end of the founding phase and single foundresses were on the nests. We used forceps to keep foundresses and usurpers 1 cm from the nest surfaces. 2002). any research about the chemical mechanisms employed by nonspecialized. Beekman & Oldroyd 2008) and intraspecific parasites have been pointed out as potential obligate preparasites or incipient obligate parasites (Taylor 1939.e. The tests lasted 1 min from the first unambiguous reactions by the offspring. P ¼ 0.336). Often. comprising the host nests and the host immature brood. 1996. . we have not come across any published research investigating the chemical ecology of intraspecific social parasites through the analysis of their hydrocarbon blends. Ruther et al. The tests were performed at least 1 day after the offspring emerged.e. 1997). Consequently. None the less. We removed their foundresses and placed them separately in glass jars.674. their mother) and usurper (i. Savolainen & Vepsalainen 2003. Bagnères & Lorenzi 2010). 1996). biglumis (Lorenzi & Turillazzi 1986). fleeing from the nest by flying. Cervo 2006). Cervo 2006. BonavitaCougourdan et al. At the same time. 2004). and ‘wait’ for the host brood to emerge. The original. 2004). We kept foundresses and usurpers on their halfcolonies for 8 days. We cut their paper combs in two with scissors so that the larger number of pupae was intact and the numbers of cells and old brood (large larvae and pupae) of the two parts were similar (foundress half-colonies: 17e28 cells. Lorenzi & Cervo 1992. we recorded the responses by 1.068).13 Æ 0. The offspring emerged after we removed foundresses or usurpers. that is. then we removed and froze them in individual glass vials (À18  C). P ¼ 0.

0 (SPSS Inc. because both peaks contained mixtures of branched and linear alkanes (in unknown proportions). We then assessed the performance of the discriminant functions on the remaining cases (usurpers and the nest paper of their half-colonies). Behavioural data came from pairs of sisters that emerged from foundress or from usurper half-colonies.13-dimethylhentriacontane) and peak 58 þ 59 (n-tetratriacontane and 3.80 Æ 1. Test for Chemical Mimicry Because our preliminary analyses showed that branched hydrocarbons played a role in the usurpers’ chemical strategies. For each extract. we analysed behavioural data as paired data by using a general linear model for repeated measures (GLM.. the usurpers would not chemically mimic their host colonies. see Garson 2008 for procedures).9-þ3. we computed the mean frequency of attacks as the sum of the behaviours recorded in 1 min in each half-colony divided by the number of adult offspring in that half-colony. biglumis are either linear or branched. they have binomial distributions and we analysed them by using a GLM with binomial errors and logit link function. Consequently.9-þ3. 0. (1) We calculated the overall amount of hydrocarbons (in ng per mg of wasp or piece of nest paper) as the overall sum of peak areas  800 ng divided by the area of the C20 peak in that extract. The DA used the colony as a grouping variable (within-group covariance matrix) and entered geomean-log-transformed data as independent variables (Mahalanobis distance method). We also measured the surplus (if any) in the amount of hydrocarbons in usurper half-nests as the difference in the overall amounts of hydrocarbons between matched pairs of usurper and foundress nest papers..15-dimethyltritriacontane) by two. for each extract. wasps weighed 31. We used half of the cases as a training subset and the remaining half as a test subset and we derived the discriminant functions on only a portion of the cases (foundresses and the nest paper of their half-colonies). Test for Chemical Transparency We tested how different from each other (if at all) the proportions were of branched to linear hydrocarbons in foundresses and usurpers. after controlling for normality). Descriptive statistics are given as means Æ 1 SE. U.S. after 1/squareroot transformation to meet assumptions of normality and homoscedasticity. We handled these compositional data (percentage of compounds) by using the log-ratio transformation (Aitchison 1982).40 mg. Because the hydrocarbons in P. unless otherwise stated. Therefore we excluded linear alkanes from the analysis. We analysed log-ratio-transformed data by using a stepwise multiple discriminant analysis (DA). we computed the natural log of the proportion of each peak and divided it by the geometric mean of the proportions of the branched alkanes.13-þ3. eight foundresses. and in the nest paper of their respective half-colonies. repeated measures. We weighed the wasps and the pieces of nest paper using a precision balance (Precisa 125A). The GC was equipped with a flame ionization detector and a Chrompack CPSIL5 WCOT CB nonpolar capillary column (25 m. / Animal Behaviour 82 (2011) 1143e1149 1145 For our analysis of behavioural data.M. Test for Chemical Insignificance We tested for differences in the overall amount of hydrocarbons between matched pairs of foundresses and usurpers and between matched pairs of foundress half-colonies and usurper half-colonies (GLM. BrayeCurtis similarities were computed in PAST (Hammer et al. We also excluded two branched-alkane peaks (peak 11a: 3-methylhexacosane. with GreenhouseeGeisser correction). we analysed the chemical profiles of 16 half-colonies. the resulting value was divided by the weight of the wasp or piece of nest paper. A few peaks consisted of a mix of coeluted hydrocarbons. 2001). If the DA misclassified usurpers and the nest paper of their half-colonies by colony. Helium was the carrier gas (1 bar). The large majority of peaks consisted of single hydrocarbons. (2) We also calculated the relative proportion of branched to total hydrocarbons as the amount of branched hydrocarbons in ng divided by the total amount of hydrocarbons in that extract. We tested whether the attacks of offspring from usurper half-colonies against usurpers increased as the distances between the usurper and her half-nest in the DA space increased (Pearson correlation. we had 51 branched-hydrocarbon peaks (chain length between 24 and 34 carbon atoms) and we computed their relative proportions.11þ3.25 mm. We used a 15 s splitless method of injection. We added to each extract 800 ng of n-C20 as an internal standard to quantify the amount of hydrocarbons. IL. On average.11-þ3. the temperature increased to 320  C at a rate of 5 /min. peak 42a: 5-methylhentriacontane) that were present in less than 25% of the samples. We expected that DA would discriminate significantly between foundresses and the nest paper of their halfcolonies by colony. 0. We extracted the hydrocarbons by dipping each wasp or piece of nest paper separately into 1 ml of pentane for 75 s. We computed the distances between usurpers and their half-nests in the DA space as the differences between the values of the discriminant function scores for usurpers and those for the nest paper of their respective half-colonies. To this end.36 Æ 2. We divided the integration area of peak 46 þ 47 (n-docotriacontane and 3. RESULTS Behavioural Tests The aggressive responses of sisters to foundresses and usurpers depended on whether the sisters had emerged from the foundress or the usurper half-colonies as shown by a significant interaction . namely. we checked whether foundresses and usurpers had profiles of branched hydrocarbons similar to those of their respective half-colonies. we checked whether the surplus in the amounts of hydrocarbons was correlated with the similarity of the chemical profiles of the usurpers and those of their respective half-colonies (as measured by BrayeCurtis similarities in the percentages of the methylbranched hydrocarbons). Chemical Analyses After the behavioural tests.21 mg and the pieces of nest paper 18. Cristina Lorenzi et al. 2 ml samples were analysed by capillary gas chromatography with a Delsi Nermag DN200 gas chromatograph (GC). The data for each extract were analysed in two ways.A. We also expected that DA would successfully classify usurpers and the nest paper of their half-colonies by colony only if the usurpers chemically mimicked their host colonies. Hydrocarbon amounts were ln transformed to meet assumptions of normality and homoscedasticity. Statistical analyses were performed in SPSS Statistics 17. repeated measures). eight usurpers and a 1 cm2 piece of the paper from their half-colonies. Assuming that the surplus was caused by the usurpers. After an isotherm of 5 min. Oven temperature increased from 70  C to 150  C at a rate of 30 /min.12 mm). 1997). Therefore. Data were registered with an Enica 31 integrator and compounds were identified by comparing their mass spectra and their retention times against those recorded earlier (Lorenzi et al. Chicago.

Test for Chemical Transparency Usurpers had significantly higher proportions of branched hydrocarbons than foundresses and. This DA classification showed that 100% of foundresses and their nest paper were correctly assigned to their colonies. Function 2. In contrast. P < 0.179). F14. Test for Chemical Insignificance The variation in the overall amount of hydrocarbons between usurper and foundress half-colonies differed between wasps and nest paper.000002. Chemical Analyses The blend of epicuticular hydrocarbons of P. power was acceptable. half-colony*target of attacks: F1. these differences were highly significant (Wald c2 1 ¼ 397:549.9% of the total variance. as shown by a significant interaction term (GLM. P ¼ 0.019.161. half-colony*sample category: F1. P ¼ 0. P ¼ 0. Figure 3. which accounted for only 5.222. 1997).035. differed significantly in mean by colony (Wilks’s l42 ¼ 0.013. repeated measures. . 1).0001. The total amount of hydrocarbons as a function of half-colony and sample category (half-colony categories: foundress and usurper half-colony.006). that is. Cristina Lorenzi et al. F7. 2). The larger the surplus of hydrocarbons on the nest paper of usurper half-colonies. only 12. Fig.857. differed significantly in mean by colony as well (Wilks’s l30 ¼ 0. a significant difference (Wald c2 1 ¼ 4421:477. biglumis foundresses.7 ¼ 6.001).7 ¼ 2. P ¼ 0. The proportion of branched alkanes as a function of the half-colony in wasps and nest paper.75 0.042.42% of the total variance. The nest paper of foundress half-colonies had on average 72% of branched hydrocarbons and that of usurper half-colonies 74% (Fig. P < 0. repeated measures. Despite large variation. Foundresses tended to be attacked less by offspring from foundress half-colonies than by their sisters from usurper half-colonies (Fig.14 ¼ 8.14 ¼ 6. step 2: Wilks’s l ¼ 0. Fig. the nest paper of usurper half-colonies had significantly larger amounts of hydrocarbons than that of foundress half-colonies (F1. larger than 0.8 0. Although only 2%. foundresses had on average 62% of branched hydrocarbons and usurpers 72%.80 only for the two-way interaction). Fig. 5000 4000 3000 2000 1000 0 Foundress half-colony Usurper half-colony Category of sample Nest Wasp Figure 2.8 ¼ 4. usurpers and their nest surfaces consisted of 70 peaks representing homologous series (C24eC34) of linear and methylbranched alkanes and confirmed previous GCeMS data (Lorenzi et al. half-colony*sample category: Wald 1 ¼ 2605:72. P ¼ 0. 4). which accounted for 93. sample categories: wasp and nest paper).001.224.332. P < 0. / Animal Behaviour 82 (2011) 1143e1149 Total amount of hydrocarbons term (GLM. 3).5% of usurpers and their nest paper were correctly assigned to their original colonies. However. P ¼ 0.00001).898.0001). In contrast. Mean frequency of attacks Æ SE by Polistes biglumis sisters that emerged from the half-colonies towards their own foundresses. P ¼ 0.6 0 Category of sample Nest Wasp Attacks by workers Usurper 6 5 0 Foundress half-colony Usurper half-colony Foundress half-colony Usurper half-colony Figure 1.1146 M. Usurpers tended to be attacked less by offspring from usurper half-colonies than by their sisters from foundress half-colonies (Fig. 3).30 ¼ 10. Matched pairs of foundresses and usurpers had roughly equivalent amounts of hydrocarbons (F1. the more similar the branched-hydrocarbon profiles of the usurpers were to those of their own half-colonies (Pearson correlation: r6 ¼ 0.190. the nest papers of usurper half-colonies had significantly larger proportions of branched hydrocarbons than those of foundress half-colonies.0001).002. the discriminant model as a whole was significant (step 1: Wilks’s l ¼ 0. 1).7 0. In their epicuticular profiles. P ¼ 0. the usurpers of their sisters or their own usurpers. 2). The total amount of hydrocarbons is expressed in ng per mg of wasp or of nest paper. This suggested that usurpers and their nest paper had chemical profiles that did not match those of 7 Target of attacks Foundress Proportion of branched hydrocarbons 0. the slope of the variation was steeper for wasps than for nest paper (as shown by a significant interaction term in the c2 GLM. The proportion of branched hydrocarbons was computed as the relative amounts of branched hydrocarbons in ng divided by the total amount of hydrocarbons.003. similarly. Function 1. P < 0. Test for Chemical Mimicry Colonies composed of foundresses and pieces of their nests could be significantly distinguished from each other by the DA (Fig.015).65 0.

Surprisingly. P ¼ 0. showing how similar the half-nests were to their foundresses or usurpers. This further suggested that usurpers were not mimicking the chemical profiles of the colonies they had usurped. the stronger the aggression by the host offspring (Pearson correlation: r6 ¼ 0. a behaviour peculiar to Polistes usurpers. The nest paper is the place where newly emerged paper wasp offspring get their information about their colony’s chemical signature. the approaching wasp is rejected. which were naïve to usurpers. the proportion of branched hydrocarbons on the nest paper was higher than that on the respective wasps. which spend time rubbing their abdomen on nest surfaces (Cervo & Turillazzi 1989. Lorenzi et al. biglumis foundresses. biglumis employ a chemical strategy of integration in host colonies. Dani et al. The emerging offspring learn the signature of their colony from the nest surfaces soon after they emerge and then use the learned template to discriminate between nestmates and strangers (Gamboa 1996. usurpers and their half-nests for colonies aeh. making them qualitatively more similar to the chemical profiles of usurpers themselves. Our behavioural results show that the offspring from usurper half-colonies. Usually. the point representing the usurper half-nest had a function 1 value <À40. 1992. Cristina Lorenzi et al. Because these offspring had no contact with their usurpers before the bioassays.839. it suggested that usurpers had partially changed the chemical profiles of their nests. 1992. were relatively tolerant towards usurpers. Instead. The deposition of hydrocarbons on host nest surfaces by usurpers changed the chemical profiles of the host nests. the original colonies. Any wasp approaching the nest is accepted by the offspring if its chemical profile matches the profile they had learned from the nest and. Lorenzi & Cervo 1992. which were the ones that the offspring that emerged from usurper nests were to learn. Cervo & Dani 1996.g. 2011). 2002. usurpers are unrelated to host foundresses (e. / Animal Behaviour 82 (2011) 1143e1149 1147 20 10 0 −10 20 10 0 Function 2 −10 20 10 0 −10 20 10 0 −10 (a) (b) (c) (d) (e) (f) Foundress (g) (h) Foundress half-nest Usurper Usurper half-nest −40 −20 0 20 40 −40 −20 0 20 40 Function 1 Figure 4.g. it is not shown in the graph. This might be caused by overmarking: foundresses marked their nests by repeatedly placing their hydrocarbon marks on the top of their previous hydrocarbon marks. In addition. 1990.). Finally. but they were neither chemically insignificant nor transparent. During nest foundation. For nest f. Usurpers probably marked host nests through stroking. usurpers had chemical profiles that were richer in methyl-branched hydrocarbons than those of the foundresses. if not. 2011) and hence they exhibit chemical profiles that do not match those of host nests. In turn. a type of scent overmarking that is common in mammals (Johnston 2005. 2004). Discriminant analysis (discriminant scores) of 51 branched hydrocarbons of P. Jordan et al. Seppä et al. These adjustments influenced the host colony signatures. therefore. Layton & Espelie 1995. 1996). relative to their hosts. Espelie et al. They were not even chemical mimics of their hosts.M. The dissimilarity between the chemical profile of usurpers and that of their host nests mediated aggression by the host offspring: the larger the dissimilarity. Van Hooser et al. Dani et al. foundresses mark the nest surfaces with their own signatures (e. Therefore. usurpers placed their . usurpers will be rejected unless they change the host nest surface chemistry in a way that biases it towards their own profiles.037). they must have learnt the usurper signatures from the usurper marks on the nest surfaces. Cervo & Lorenzi 1996. DISCUSSION Facultative social parasites of P. usurpers deposited large quantities of hydrocarbons on the surfaces of their host nests.

hydrocarbons increased in quantity on the nest paper. 62. A. C. M. Annual Review of Entomology. A. Dani. 2010. Bonavita-Cougourdan. 889e892. R. Our results show that the usurpers’ chemical strategies of integration are very different from those of obligate social parasites in that usurpers mark.g. The statistical analysis of compositional data.. A.-G. 1e10. that is. by S. J. Lenoir et al. Nest exchange experiments in Polistes gallicus (L. J. C. Chemical Senses. however. Usurpers and their host nests were particularly rich in methyl-branched hydrocarbons. 327e335. Chemical deception/mimicry using cuticular hydrocarbons. 121e162. J. Journal of Entomological Science. Cambridge: Cambridge University Press. 19e37. Science. S. 2007) and therefore exhibit a preadaptation to obligate parasitism that P. 1992. Chemical taxonomy with hydrocarbons. Queller. Dani et al.U. 1988. & Jones. 44.). (Hymenoptera. de L. & Davies. 2005. D’Ettorre. & Turillazzi. 11. For example. Deciphering the recognition signature within the cuticular chemical profile of paper wasps. Social parasitism among ants: a review (Hymenoptera: Formicidae). C. S. R. & Oldroyd.. 1994. Cervo. rather than only the mark on the top. S. Insect Social Life. 1982. Bagnères). Lorenzi. Bagnères. Cini. In: Natural History and Evolution of Paper-Wasps (Ed. Bagnères et al. Vespidae). & Turillazzi. Biochemistry and Chemical Ecology (Ed. R. Most of the approximately 200 species of paper wasps behave as facultative parasites but only three monophyletic species behave as obligate parasites (Choudhary et al. pp. Then. D. Branched hydrocarbons seem to be more easily detectable and seem to convey more information than linear alkanes (Châline et al. 51e58. they virtually ‘bury’ the legitimate queen’s chemical signature to make it easier for them to trick emerging host offspring about the queen’s identity. Insectes Sociaux. & Dani. 185e193.. 83.-L. matters in host nest invasion by a wasp social parasite. However. M. Clément. Abdomen stroking behaviour and its possible functions in Polistes dominulus (Christ) (Hymenoptera. what advantage do usurpers gain by emphasizing their hydrocarbon profiles and enriching their branched-hydrocarbon components disproportionately? If branched hydrocarbons play an essential role in encoding information about a queen’s identity. we analysed a mixture of past and recent marks..-G. Poggi. C. In: Insect Hydrocarbons: Biology. C. facultative parasites may use chemical strategies of integration into host colonies that are different from those of obligate parasites. B. Cambridge: Cambridge University Press. Cristina Lorenzi et al.. Cervo. 2011). Learning and discrimination of individual cuticular hydrocarbons by honeybees (Apis mellifera). G. Myrmecological News. Châline. camouflage. Sandoz. 1994. 139e177. 30. Apart from flexibility. whose chemical signature is highly flexible and changes with the phases of host colony invasion (e. 2010. Turillazzi & M. Annual Review of Entomology. Animal Behaviour. We found a significant increase in branched hydrocarbons in both usurpers and in usurper nest paper surfaces. Polistes wasps and their social parasites: an overview. they reduce the waterproofing effect of epicuticular hydrocarbon layers (Le Conte & Hefetz 2008. 2010. 2001). R. Cambridge: Cambridge University Press. they resemble obligate social parasites.. Beekman. N. Therefore these results suggest that foundresses have a flexible chemical signature. Gibbs & Rajpurohit 2010). Blomquist. F.. 2008b).I. C. by G. 22. Destri. usurpers are not a subset of the wasp population. A. rather than mimic. 104e112. 28. biglumis usurpers are not likely to be incipient obligate parasites. Akino 2008. then successful usurpers must cover the legitimate queen’s signature with their own. M. R. We also thank Laura Azzani and two anonymous referees for their helpful comments on the manuscript. Journal of the Royal Statistical Society B. Oxford: Oxford University Press. S. F. Ratnieks.. Martin et al. / Animal Behaviour 82 (2011) 1143e1149 hydrocarbon marks on top of the hydrocarbon marks of the foundresses. Dani. N. M. 1996. Biochemistry and Chemical Ecology (Ed. Cervo. In effect. R. Cervo & Dani 1996). Behavioural Processes. Bagnères). Fight or fool? Physical strength. 2. 2011. 219e235. & Bagnères. & Liepert. such as P. R. 630e632. & Wicker-Thomas. Myrmecological News. foundresses shift to usurping conspecific nests as a conditional strategy when their colonies are killed by predators (Lorenzi & Cervo 1995). & Turillazzi. This suggests that P. 447e492. Chemical usurpation of a nest by paper wasp parasites. 2001). biglumis usurpers do not have. but we also know that some classes of compounds. Bagnères & Lorenzi 2010).-G. 2001. biglumis. What obligate parasites do next is to steal into the host social structures and mimic their hosts chemically. 272. L. Egg mimicry by cuckoos Cuculus canorus in relation to discrimination by hosts. . 1139e1145. This may explain the mismatch between the proportion of hydrocarbons of wasps and nests. (ex 60%) to M. Dusticier. 165e171. A. T. C. S. 1987. G. 43.-G. Cervo.-G. Polistes nimphus intraspecific usurpers seem to mimic host colonies (Lorenzi et al. 335. Blomquist & A. Ethology Ecology and Evolution. 2010. E. W. Bagnères). 2005. G. reviewed in D’Ettorre & Errard 1998. Behaviour in usurpers and joiners of Polistes biglumis bimaculatus (Hymenoptera Vespidae). Pheromones in social wasps. J. As a consequence. S.C. In fact. Bagnères. In fact. L. F. and phytomimesis by ants (Hymenoptera: Formicidae) and other arthropods. Chemical disguise during colony founding in the dulotic ant Polyergus rufescens Latr. 2009. This does not imply that obligate parasitism did not originate from conspecific usurpation in other species. J. & Lorenzi. The different marks probably had different proportions of linear and branched hydrocarbons because the proportions of branched and linear hydrocarbons on the wasp cuticle varied. Blomquist & A. 12. A. R.L. propaganda. C. 255e266. WestEberhard). Vitamins and Hormones. L. P. 1996. Akino. Chemical strategies to deal with ants: a review of mimicry. We are far from deciphering the communication codes encrypted in the hydrocarbon profiles of social insects. R. H. which they adjust depending on the conditions they are facing. 2008. may play key roles in recognition (Bonavita-Cougourdan et al. Blomquist & A. methyl-branched hydrocarbons. Bruschini. Brooke. R. M. Clément. 1996. When we extracted the hydrocarbons from the nest paper. R. Strassmann. Martin. Cervo. In nature. pp. B. 1998. When obligate social parasites enter host nests and face their hosts for the first time. In: Insect Hydrocarbons: Biology. Funding was provided by M.. Nature. Proceedings of the Royal Society B. 2007. A. 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