Long-Lasting Insecticide-Treated Netting Affects Reproductive Output and Mating Behavior in Tribolium castaneum (Coleoptera: Tenebrionidae) and Trogoderma variabile (Coleoptera: Dermestidae).

Preventing insect infestations is a critical component for establishing a pest management plan for stored-product insects. Long-lasting insecticide-treated netting (LLIN) is a potential tool to reduce insect movement by providing a chemical barrier, where insects may be able to physically pass through but ultimately die after exposure to the netting. Sublethal effects, such as reduced movement immediately after exposure and reduced ability to colonize, have been reported. Here we examine the sublethal effects of exposure to LLIN on two beetle species, Trogoderma variabile Ballion, warehouse beetle, and Tribolium castaneum Herbst, red flour beetle. We found that both female and male T. castaneum exposed to LLIN produced significantly less adult progeny than those exposed to untreated netting. Adult progeny output did not differ for T. variabile, but survivorship increased in T. variabile females exposed to LLIN. Importantly, the overall net reproductive rate was significantly decreased for both T. variabile and T. castaneum. The number of copulation attempts did not differ between males or females exposed to LLIN compared to untreated netting, but males exposed to LLIN showed increased durations of attempted and successful copulation events. This research demonstrates that the implications of LLIN exposure extend past direct mortality, with sublethal effects on reproductive output potentially increasing the effectiveness of this tool for preventing insect infestations.

Callosobruchus embryo struggle to guarantee progeny production.

We conducted a series of experiments to test insect embryo capability to survive and increase reproductive investment during early development after short exposure to essential oils. We used Callosobruchus maculatus as a model insect and eucalyptus leaf and flower essential oils. Both essential oils exhibited toxicity against C. maculatus embryos and adults. However, flower essential oil was more toxic. A fetus exposed to essential oils tried to make the best of a bad situation and compensate essential oils harmful effects in the later life stages. Insect progeny production guarantee resulted in a trade-off between reproduction and female longevity. The insect also could alter fitness and reproductive behavior including, mating latency reduction, copulation duration increase, and copulation success rate raise in adulthood. Flower essential oil-exposed embryos were more successful in increasing copulation duration, and leaf essential oil-exposed embryos achieved more copulation success and less mating latency. These consequences persisted until F1 generation that was not directly exposed to essential oil. However, the F2 generation could concur with the harmful effects of essential oils. C. maculatus embryo might use epigenetic mechanisms to guarantee progeny production. Reproductive behavior changes and the trade-off can be evolutionary mechanisms to save species from possible extinction in deleterious situations.

The effects of 28-day early-life exposure to triphenyl phosphate (TPhP) on odor preference and sexual behavior in female rats.

Many chemical substances are detectable in house dust, and they are consequently taken into our bodies via the mouth and nose. Triphenyl phosphate (TPhP), a flame retardant that has an estrogen-like effect in vitro, is present in house dust at high concentrations. Estrogen exposure during development has significant influences on reproductive behavior in rodents, and its effects persist until maturity. In the present study, we investigated the effect of early life exposure to TPhP on the reproductive behavior of female rats. Oral treatment with TPhP (25 or 250 mg/kg), ethinyl estradiol (EE; 15 µg/kg) as a positive control, or sesame oil as a negative control, were given to female rats (from birth to 28 days of age). The 8-week-old rats were bilaterally ovariectomized. At 12-15 weeks of age, the rats were subjected to odor preference and sexual behavior tests. In the odor preference test, the oil group showed significantly higher preference for male odor than female odor, but the low-dose TPhP treatment group lost the preference for male odor, indicating a possible outcome of early life TPhP exposure on sexual recognition. In the sexual behavior test, both the EE and TPhP treatment groups displayed significantly less proceptive behavior. These results suggest that early life exposure to TPhP disturbs the normal sexual behavior of female rats.

Comparison of swarming, mating performance and longevity of males Anopheles coluzzii between individuals fed with different natural fruit juices in laboratory and semi-field conditions.

BACKGROUND: It is assumed that malaria vectors feed on locally available nectar sources to obtain energy. Sugar feeding is energetically critical for the Anopheles male swarming and mating activities. However, little is known about the impact of local nectar feeding on male physiological development and its consequences on male mosquito life traits in the malaria control context. This study aimed to evaluate the influence of local fruit juices on the life traits of males Anopheles coluzzii. METHODS: Swarming characteristics (number of males in swarm, number of mating pairs, and swarm duration) in semi-field conditions; mating rate and longevity in a laboratory setting were compared between males An. coluzzii fed exclusively with mango, papaya or banana juices. The trophic preference was investigated in semi-field conditions. RESULTS: The results of this study showed that in the laboratory, mosquitoes fed with papaya juices lived on average longer (10 days) than those fed with banana or mango juices (5 days) and had higher a mating rate (53%) than those fed with banana juice (40%). In the semi-field, the swarm size of mosquitoes fed with banana juice (85 males) was larger than that of mosquitoes fed with mango juice (60 males). The number of mating pairs formed from banana-fed male swarms (17 mating pairs) was higher than that formed from mango-fed male swarm (8 mating pairs). There was no difference in swarming duration between male treatments. Male mosquitoes had a preference for papaya and banana juices. CONCLUSIONS: The results indicate that the origin of plant-derived feeding is an important factor in the survival and reproduction of mosquitoes. This calls for further investigations of chemical contents of nectars and their impact on the physiological development of mosquitoes.

Differential sensitivity of alcohol drinking and partner preference to a CRFR1 antagonist in prairie voles and mice.

Available pharmacotherapies to treat alcohol use disorder (AUD) show limited efficacy. Preclinical studies in mice and rats suggested that antagonists of the corticotropin releasing factor receptor 1 (CRFR1) could be more efficacious for such treatment. However, clinical trials with CRFR1 antagonists were not successful. While a number of potential explanations for this translational failure have been suggested, we hypothesized that the lack of success in clinical trials could be in part due to different neuroanatomical organization of the CRFR1 system in mice and rats versus humans. The CRF system in prairie voles (Microtus ochrogaster), a socially monogamous rodent species, also shows differences in organization from mice and rats. To test our hypothesis, we compared the efficacy of a potent CRFR1 antagonist, CP-376,395, to modulate alcohol drinking in male and female prairie voles versus male and female C57BL/6J mice using an almost identical 2-bottle choice drinking procedure. CP-376,375 (10 and 20 mg/kg, i.p.) significantly decreased alcohol intake (but not alcohol preference) in mice, but not prairie voles. Furthermore, administration of this antagonist (20 mg/kg, i.p.) prior to the partner preference test (PPT) decreased partner preference (PP) in male prairie voles. These findings support our hypothesis that the greater efficacy of CRFR1 antagonists to suppress alcohol consumption in mice and rats versus other mammalian species could be due to the differences in organization of the CRFR1 system between species. They further indicate that activity of the CRFR1 system is necessary for the formation of pair-bonds, but not consumption of high doses of alcohol. Overall, we suggest that testing potential pharmacotherapies should not rely only on studies in mice and rats.

Early-exposure to new sex pheromone blends alters mate preference in female butterflies and in their offspring.

While the diversity of sex pheromone communication systems across insects is well documented, the mechanisms that lead to such diversity are not well understood. Sex pheromones constitute a species-specific system of sexual communication that reinforces interspecific reproductive isolation. When odor blends evolve, the efficacy of male-female communication becomes compromised, unless preference for novel blends also evolves. We explore odor learning as a possible mechanism leading to changes in sex pheromone preferences in the butterfly Bicyclus anynana. Our experiments reveal mating patterns suggesting that mating bias for new blends can develop following a short learning experience, and that this maternal experience impacts the mating outcome of offspring without further exposure. We propose that odor learning can be a key factor in the evolution of sex pheromone blend recognition and in chemosensory speciation.

Wild bacterial probiotics fed to larvae of mass-reared Queensland fruit fly [Bactrocera tryoni (Froggatt)] do not impact long-term survival, mate selection, or locomotor activity.

Queensland fruit fly [Bactrocera tryoni (Froggatt), Diptera, Tephritidae] is the most devastating insect pest impacting Australian horticulture. The Sterile Insect Technique (SIT) is an important component of tephritid pest management programs. However, mass-rearing and irradiation (to render insects sterile) may reduce the fitness and performance of the insect, including the ability of sterile males to successfully compete for wild females. Manipulation of the gut microbiome, including the supplementation with bacterial probiotics shows promise for enhancing the quality of mass-reared sterile flies, however there are fewer published studies targeting the larval stage. In this study, we supplemented the larval stage of mass-reared B. tryoni with bacterial probiotics. We tested several individual bacteria that had been previously isolated and characterized from the gut of wild B. tryoni larvae including Asaia sp., Enterobacter sp., Lactobacillus sp., Leuconostoc sp. We also tested a consortium of all four of these bacterial isolates. The fitness parameters tested included adult survival in field cages, laboratory mate selection of bacteria supplemented males by bacteria nonsupplemented females, and laboratory locomotor activity of adult flies. None of the bacterial probiotic treatments in the current study was significantly different to the control for field survival, mate selection or locomotor activity of adult B. tryoni, which agree with some of the other studies regarding bacterial probiotics fed to the larval stage of tephritids. Future work is needed to determine if feeding the same, and/or other probiotics to adults, as opposed to larvae can positively impact survival, mating performance, mating competitiveness and locomotor activity of B. tryoni. The bacterial group(s) and function of bacterial species that increase fitness and competitiveness is also of interest to tephritid mass-rearing programs.

Glucocorticoids, male sexual signals, and mate choice by females: Implications for sexual selection.

We review work relating glucocorticoids (GCs), male sexual signals, and mate choice by females to understand the potential for GCs to modulate the expression of sexually selected traits and how sexual selection potentially feeds back on GC regulation. Our review reveals that the relationship between GC concentrations and the quality of male sexual traits is mixed, regardless of whether studies focused on structural traits (e.g., coloration) or behavioral traits (e.g., vocalizations) or were examined in developmental or activational frameworks. In contrast, the few mate choice experiments that have been done consistently show that females prefer males with low GCs, suggesting that mate choice by females favors males that maintain low levels of GCs. We point out, however, that just as sexual selection can drive the evolution of diverse reproductive strategies, it may also promote diversity in GC regulation. We then shift the focus to females where we highlight evidence indicating that stressors or high GCs can dampen female sexual proceptivity and the strength of preferences for male courtship signals. Hence, even in cases where GCs are tightly coupled with male sexual signals, the strength of sexual selection on aspects of GC physiology can vary depending on the endocrine status of females. Studies examining how GCs relate to sexual selection may shed light on how variation in stress physiology, sexual signals, and mate choice are maintained in natural populations and may be important in understanding context-dependent relationships between GC regulation and fitness.

Mate choice in a polluted world: consequences for individuals, populations and communities.

Pollution (e.g. by chemicals, noise, light, heat) is an insidious consequence of anthropogenic activity that affects environments worldwide. Exposure of wildlife to pollutants has the capacity to adversely affect animal communication and behaviour across a wide range of sensory modalities-by not only impacting the signalling environment, but also the way in which animals produce, perceive and interpret signals and cues. Such disturbances, particularly when it comes to sex, can drastically alter fitness. Here, we consider how pollutants disrupt communication and behaviour during mate choice, and the ecological and evolutionary changes such disturbances can engender. We explain how the different stages of mate choice can be affected by pollution, from encountering mates to the final choice, and how changes to these stages can influence individual fitness, population dynamics and community structure. We end with discussing how an understanding of these disturbances can help inform better conservation and management practices and highlight important considerations and avenues for future research. This article is part of the theme issue 'Linking behaviour to dynamics of populations and communities: application of novel approaches in behavioural ecology to conservation'.

Advertisement of unreceptivity - Perfume modifications of mason bee females (Osmia bicornis and O. cornuta) and a non-existing antiaphrodisiac.

Females of many monandrous insect species announce their receptivity either by specialised sex-pheromones or by a signature mixture of cuticular hydrocarbons (CHCs). The trigger that shuts down the sex-pheromone release or initialises a change in CHC bouquet is thought to be either the mating per se or male pheromones transferred during copulation. Besides a conversion of female volatiles, the application of antiaphrodisiacs, male derived pheromones that render mated females unattractive to competitors, is another strategy to protect females from further sexual chasings. This simple pattern becomes more complicated in the monandrous mason bees Osmia bicornis (syn: O. rufa) and O. cornuta due to a post-copulation phase in their mating sequence. Males display a stereotypic behaviour right after the intromission that induces females' unreceptivity. This post-copulatory display is predestined both to trigger a transition of the CHC profile and for the application of an antiaphrodisiac. However, the postulated antiaphrodisiac was not detectable even on freshly mated females. Moreover, the male's post-copulatory display did not trigger a change in the CHC bouquet and neither did the insemination. Instead the CHC profile of freshly emerged females changes into the bouquet of nesting females simply by age as an ontogenetic process in both Osmia species. This autonomous change in the CHC profile coincides with an age-specific decrease of young female's willingness to mate. How the resulting short period of female receptivity without back coupling by storage of sperm and the lack of an antiaphrodisiac fit into the behavioural ecology of the studied mason bee species is discussed.

Corticosterone mediated mate choice affects female mating reluctance and reproductive success.

The study of stress-related hormones as mediators of sexual selection has traditionally focused on the effect of glucocorticoids on male quality and competing ability. However, environmental stressors are expected to affect both males and females, and the strength of sexual selection might be affected by changes in female mating decisions, a hypothesis that has rarely been tested. Here, we investigated whether female common lizard (Zootoca vivipara) mating behaviour and mating preferences are affected by different levels of administered corticosterone and conditioned by the familiarity of their partners, which is known to influence Z. vivipara social behaviour. To this end, two females, one corticosterone-treated and one control female, were simultaneously presented with an unfamiliar male and the following day with either a familiar or an unfamiliar male. Females treated with corticosterone (Cort) were more aggressive towards males and mated less. Furthermore, copulation probability in Cort females, but not in control females, increased with body size. On the second day, Cort females only mated with familiar partners. In contrast, male behaviour towards females was not affected by treatment and only bigger males successfully copulated with Cort females. This shows that corticosterone directly affected female mating behaviour and mating preferences, while male mating behaviour was unaffected by the female's level of corticosterone. Environmental and social stressors may affect reproductive strategies of females, the strength of sexual selection, and sexual conflict through their effects on female glucocorticoid levels, potentially in a wide range of species.

Mating under the influence: male Siamese fighting fish prefer EE2-exposed females.

Countless pharmaceuticals and endocrine disrupting chemicals (EDCs) exist on the market with more added each day. Many of these compounds are not removed during the wastewater treatment process and enter bodies of water in their active form. EDCs are known to have physiological and behavioral effects in exposed organisms. Exposure to the synthetic estrogen 17α-ethinylestradiol (EE2), a common EDC found in birth control pills, has been found to lead to population collapse after only a few generations in some fish species. Mechanisms identified as potential driving forces for collapse include feminization of males and altered fecundity in both sexes. However, an additional way in which EE2 could lead to population collapse is by altering courtship behavior, which could then change mating preferences and decrease mating opportunities. The current study had the following objectives: determine if exposing female Siamese fighting fish, Betta splendens, to EE2 changes mate choice in males; assess if the dose and duration of female exposure matters; and examine if exposing males to EE2 influences their mating preferences. Both unexposed and exposed males were presented with pairs of females that differed in EE2 dose and exposure duration. The results indicate that males were more responsive to EE2-exposed females than unexposed females, with males being most responsive to females exposed to the low versus high dose. Furthermore, exposed males responded less overall than unexposed males. If EE2 concentration increases in the environment, the likelihood of successful mating could decrease and, therefore, potentially lead to adverse population impacts.

The endocrine disruptor, 17α-ethinyl estradiol, alters male mate choice in a freshwater fish.

Among the handful of studies on the behavioural effects of endocrine disrupting chemicals (EDCs), only a few have set out to disentangle the mechanisms underpinning behavioural changes. In fish, previous studies have shown that both visual and chemical cues play an important role in mate choice. As such, contaminant-induced changes in either transmission or perception of mate choice cues could have direct implications for individual's fitness. One widespread contaminant of environmental concern is 17α-ethinyl estradiol (EE2), a synthetic estrogen used in the contraceptive pill. Here, we investigated the impacts of EE2 exposure (28 days; measured concentration 14 ng/L) on visual and chemical communication in wild guppies (Poecilia reticulata). Using a standard dichotomous mate choice assay, we first gave individual males (either control or EE2-exposed) the opportunity to court two size-matched females (one control and one EE2-exposed) using only visual cues. We then introduced chemical cues of females (control and EE2-exposed) to the trial tank. We found that there was no significant effect of EE2-treatment on total time males spent associating with the females, when given only visual cues. There was, however, a significant effect on male courtship behaviour, with both control and EE2-exposed males spending more time performing 'sigmoid' displays towards the visual cues of control females compared to EE2-exposed females. When males were presented with both visual and chemical female cues simultaneously, we found that males spent more time courting control females that were paired with EE2-chemical cues. Not only does our study uncover a previously unknown behavioural impact of EE2-exposure on chemical cues, but demonstrates that EE2-exposure can exert complex effects on visual and chemical communication in a mate choice context. Finally, we contribute to the discussion of intraspecific variability by providing data on the potential trade-offs underpinning contaminant-induced behavioural changes.

Body Odor and Sex: Do Cuticular Hydrocarbons Facilitate Sexual Attraction in the Small Hairy Maggot Blowfly?

Cuticular hydrocarbons (CHCs) play an important role as contact pheromones in insects, particularly in flies. However, for many fly taxa our understanding of the importance of CHCs in sexual communication is limited. Within the family Calliphoridae (blowflies), sex-specific differences in CHCs have been reported for several species, but there is no evidence that CHCs facilitate sexual behavior. In order to elucidate the function of CHCs in Calliphoridae, studies combining behavioral and chemical analyses are required. The present study used gas chromatography/mass spectrometry, along with behavioral assays, to assess whether CHCs facilitate sexual attraction in the small hairy maggot blowfly, Chrysomya varipes. The specific aims were to: 1) determine if CHCs differ between the sexes and 2) assess whether flies exhibit positive chemotaxis to CHCs of the opposite sex. Fifty-two hydrocarbons common to both sexes were identified, and quantitative differences for numerous CHCs were observed between the sexes. However, behavioral assays provided no evidence that flies were attracted to CHCs of the opposite sex, challenging the hypothesis that CHCs facilitate sexual attraction in Ch. varipes. In contrast to other blowflies, Ch. varipes males invest heavily in courtship displays and ornamentation, so we speculate that visual communication in this species may have relaxed sexual selection for chemical communication. More broadly, our findings support suggestions that CHCs may not always facilitate insect sexual communication.

Female assortative mate choice functionally validates synthesized male odours of evolving stickleback river-lake ecotypes.

During mate choice decisions, females of many vertebrates use male olfactory cues to achieve immunogenetic optimality of their offspring. Three-spined sticklebacks ( Gasterosteus aculeatus) populating habitats that differ in their parasite communities evolve locally adapted combinations of genetic variants encoded at the major histocompatibility complex (MHC). Such adaptation confers optimal resistance to the local parasite fauna. Immunogenetic signatures co-evolved with local parasites favour population-specific assortative mate choice behaviour. Previous studies have shown that female sticklebacks evaluate male MHC-associated olfactory cues during the process of mate choice, but how habitat-specific information is exchanged between males and females has remained elusive. Here, we directly demonstrate the molecular nature of the olfactory cue providing habitat-specific information. Under controlled laboratory conditions, females that are ready to mate prefer mixtures of synthetic MHC peptide ligands mimicking the optimal allele number of their original population. These results imply that female sticklebacks can determine the number of MHC alleles of their prospective mates, compare it to their own immunogenetic status, and, if optimal with respect to the immunogenetic complementarity, accept the male as mate. Our results suggest a potentially common mechanism of ecological speciation in vertebrates that is based on the olfactory assessment of habitat-specific immunogenetic diversity.

Mate choice could be random in female rats (Rattus norvegicus).

Female mate choice is often investigated in terms of reproductive success in order to understand how male characteristics contribute to sexual attractiveness. Previous studies have found that females rats prefer mating with their first encounter rather than males visited subsequently, suggesting that the rewarding value of this first encounter is enough to reinforce mating with the first partner. Using a multiple chambers paradigm, we allowed female rats to copulate freely with three males placed each in a different chamber. Then, we switched the males' position, and let the female interact with them freely again within the same session. We tested whether female mate choice was relying rather on a preferred male rat or on a preferred mating location. The results showed that females spent most time with the male in the chamber of 1st entry in the beginning, but as soon as male rats switched chambers, the female rat continued to copulate with the new male in the same chamber of 1st entry, instead of mating with her previously preferred male rat. This suggests that the male preference is an artefact of location preference. Therefore, female mate choice seems to be rather random than the consequence of an individual choice based on male characteristics. This finding, although contradictory with the intuitive feeling that mate choice is a crucial feature in sexual and reproductive behavior, is supported by several recent observations. In the coming years, behavioral neuroscience should bring light to the brain processes at work in random mate choice.

Olfactory conditioned same-sex partner preference in female rats: Role of ovarian hormones.

The dopamine D2-type receptor agonist quinpirole (QNP) facilitates the development of conditioned same-sex partner preference in males during cohabitation, but not in ovariectomized (OVX) females, primed with estradiol benzoate (EB) and progesterone (P). Herein we tested the effects of QNP on OVX, EB-only primed females. Females received a systemic injection (every four days) of either saline (Saline-conditioned) or QNP (QNP-conditioned) and then cohabited for 24h with lemon-scented stimulus females (CS+), during three trials. In test 1 (female-female) preference was QNP-free, and females chose between the CS+ female and a novel female. In test 2 (male-female) they chose between the CS+ female and a sexually experienced male. In test 1 Saline-conditioned females displayed more hops & darts towards the novel female, but QNP-conditioned females displayed more sexual solicitations towards the CS+ female. In test 2 Saline-conditioned females displayed a clear preference for the male, whereas QNP-conditioned females displayed what we considered a bisexual preference. We discuss the effect of dopamine and ovarian hormones on the development of olfactory conditioned same-sex preference in females.

Secondary compounds from exotic tree plantations change female mating preferences in the palmate newt (Lissotriton helveticus).

Selection can favour phenotypic plasticity in mate choice in response to environmental factors that alter the costs and benefits of being choosy, or of choosing specific mates. Human-induced environmental change could alter sexual selection by affecting the costs of mate choice, or by impairing the ability of individuals to identify preferred mates. For example, variation in mate choice could be driven by environmentally induced differences in body condition (e.g. health) that change the cost of choosiness, or by environmental effects on the ability to detect or discriminate sexual signals. We teased apart these possibilities experimentally, by comparing female mate choice in the palmate newt Lissotriton helveticus between environments that mimic water from either native oak forests or exotic eucalypt plantations. In laboratory two-choice mate trials in clean water, females with prolonged exposure (21 days) to waterborne chemicals leached from eucalypt leaves did not preferentially associate with the male with a stronger immune response, but females exposed to water with chemicals from oak leaves did. In contrast, female choice was unaffected by the immediate presence or absence of eucalypt leachates during mate choice (using only females previously held in oak-treated water). The habitat-related change in female choice we observed is likely to be driven by effects of eucalypt leachates on female physiology, rather than immediate inhibition of pheromone transmission or blocking of pheromone reception.

Prepubertal ovariectomy modulates paced mating behavior but not sexual preference or conditioned place preference for mating in female rats.

The present study investigated whether the presence or absence of peripubertal ovarian hormones affects sexual preference and conditioned place preference for paced mating in adult female rats primed with 10µg estradiol benzoate and 1mg progesterone. Ovariectomy (OVX) occurred either before or after pubertal development, and 4weeks later rats began a series of behavioral tests. Rats with ovaries removed before the pubertal timeframe (Prepubertal OVX) were more active, more likely to withdrawal from the male compartment, and did not discriminate between mounts and intromissions during paced mating relative to rats with ovaries during puberty (Adult OVX). Both Adult OVX and Prepubertal OVX rats showed a higher preference for the male when hormone primed vs. oil treated and a conditioned place preference for paced mating behavior. The results of the present study demonstrate that some, but not all, aspects of female sexual behavior require ovarian hormones during puberty.

Heterosexual experience prevents the development of conditioned same-sex partner preference in male rats.

Sexual partner preferences can be strengthened, weakened or even drastically modified via Pavlovian conditioning. For example, conditioned same-sex partner preference develops in sexually-naïve male rats that undergo same-sex cohabitation under the effects of quinpirole (QNP, D2 agonist). Here, we assessed the effect of prior heterosexual experience on the probability to develop a conditioned same-sex preference. Naïve or Sexually-experienced males received either Saline or QNP and cohabited during 24h with a male partner that bore almond scent on the back as conditioned stimulus. This was repeated every 4days for a total of three trials and resulted in four groups (Saline-naïve, Saline-experienced, QNP-naïve, QNP-experienced). Social and sexual preference were assessed four days after the last conditioning trial in a drug-free test in which experimental males chose between the scented familiar male and a novel sexually receptive female. Results showed that Saline-naïve, Saline-experienced and QNP-experienced displayed a clear preference for the female (opposite-sex). By contrast, only QNP-naïve males displayed a same-sex preference. Accordingly, QNP-experienced males were not affected by the conditioning process and continued to prefer females. We discuss the effects of copulation and D2 agonists on the facilitation and/or disruption of conditioned partner preferences.