SUMMARYFemales are often thought to use several cues and more than one modality in selection of a mate, possibly because they offer complementary information on a mate's suitability. In the red mason bee, Osmia rufa, we investigated the criteria a female uses to choose a mating partner. We hypothesized that the female uses male thorax vibrations and size as signs of male viability and male odor for kin discrimination and assessment of genetic relatedness. We therefore compared males that had been accepted by a female for copulation with those rejected, in terms of their size, their immediate precopulatory vibrations (using laser vibrometry), the genetic relatedness of unmated and mated pairs (using microsatellite markers) and emitted volatiles (using chemical analyses). Females showed a preference for intermediate-sized males that were slightly larger than the modal male size. Furthermore, male precopulatory vibration burst duration was significantly longer in males accepted for copulation compared with rejected males. Vibrations may indicate vigor and assure that males selected by females are metabolically active and healthy. Females preferentially copulated with males that were genetically more closely related, possibly to avoid outbreeding depression. Volatiles of the cuticular surface differed significantly between accepted and rejected males in the relative amounts of certain hydrocarbons, although the relationship between male odor and female preference was complex. Females may therefore also use differences in odor bouquet to select among males. Our investigations show that O. rufa females appear to use multiple cues in selecting a male. Future investigations are needed to demonstrate whether odor plays a role in kin recognition and how the multiple cues are integrated in mate choice by females. Supplementary material available online at

Climate change and the resulting changes in air temperature are known to have a major influence on most animals, especially poikilothermic insects, because they depend on the high enough temperatures to function. Previous studies have shown that various signals can be affected by changes in temperature. However, research into the effect of temperature on mating signals and subsequently communication between mates and on female choice is still rare. In the red mason bee, Osmia bicornis, which emerges early in spring and is therefore subject to extensive temperature changes, females choose suitable males based on, among other criteria, their thorax vibrations and odor. So far there has been no research into the effect of temperature changes on these signals. We therefore investigated whether the environmental temperature has an influence on the male's mating signals by measuring vibrations using a laser vibrometer and collecting male and female odor at different temperatures. Furthermore, we performed behavioral experiments in order to show whether there is an effect of temperature‐induced changes and female choice. Our results showed that rejected males differed in their vibrations between the two temperature settings but accepted ones did not. Temperature changes therefore seem to have a stronger effect on those males that are rejected by the females, whereas the accepted males are the ones that can produce desirable signals despite temperature fluctuations. Furthermore, we found that the differences in odor profiles were greater between temperature settings than between the sexes and that females change their preference for odor with temperature. We conclude that temperature strongly influences the male mating signals and therefore may have a major impact on sexual selection in this species. This is an important aspect to consider, not only in future studies on mating behavior, but also in view of our ever raising temperatures.

Differences in female preference for certain male characteristics can be a driving force for population divergence and speciation [1-4]. During precopulation, females of the red mason bee, Osmia bicornis, choose suitable males based on, among other criteria, their thoracic vibrations [5]. These vibrations are thought to be a signal of a male's fitness with females choosing the strongest males that can vibrate for the longest time [5]. The precise role of such vibrational signals, however, has not been determined by bioassays, and the vibrations might also play a role in species recognition [6]. There are two main subspecies of O. bicornis in Europe distinguishable only by a single morphological trait [7] (Figure S1). We therefore developed a new bioassay allowing us to impose the vibrations of one live male onto another in order to discern possible selective mate choice by females from O. bicornis originating from different regions of Europe. Females showed strong preference for males from their own region, and male vibrations were the main signal involved in this choice. Thus, vibrational signals encode not only fitness but also information about the region of origin indicating that divergence exists between the different European O. bicornis populations, which might ultimately lead to speciation. These results provide new insights into the scope of vibrational communication in bees, a group previously considered to rely predominantly on chemical signals [8, 9]. Our newly developed method should shed further light on many exciting questions concerning vibrational communication in bees and other animal taxa.

1. Recent reports on bee health suggest that sublethal doses of pesticides have negative effects on wild bee reproduction and ultimately on their population growth.2. Females of the solitary horned mason bee Osmia cornuta, evaluate thoracic vibrations and odours of males to assess male quality. When certain criteria are met, the female accepts the male and copulates. However, these signals were found to be modified by sublethal doses of pesticides in other hymenopterans. Here, we tested whether sublethal doses of a commonly used fungicide (Fenbuconazole) impact male quality signals and mating success in O. cornuta. Males exposed to fenbuconazole exhibited reduced thoracic vibrations and an al-tered cuticular hydrocarbon profile compared to the control bees. Moreover, males exposed to the fungicide were less successful in mating than control males. Synthesis and applications.Our results indicate that a low toxicity fungicide can negatively affect male reproductive success by altering behavioural and chemical cues. This could explain the decreasing pollinator populations in a pesticidepolluted environment. This study highlights the need for a more comprehensive approach, including behaviour and chemical cues, when testing new pesticides and a more cautionary approach to the pesticides already used on crops.