Size-Dependent Mating Success in Pink Bollworm moths: Insights into Mating Disruption Effects

Size-Dependent Mating Success in Pink Bollworm moths: Insights into Mating Disruption Effects | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Size-Dependent Mating Success in Pink Bollworm moths: Insights into Mating Disruption Effects Shevy Waner Rips, Michal Motro, Uzi Motro, Oren Kolodny, Ally Harari This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7842351/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Mating disruption is an environmentally friendly pest management technique that interferes with pheromone communication in moths, disrupting males’ ability to locate females, thereby reducing pest populations in agricultural systems. This study explores how mating disruption affects mating success and size distribution in the pink bollworm ( Pectinophora gossypiella ), a major cotton pest worldwide. The species pheromone normally consists of a 1:1 ratio of Z,Z- and Z, E-7,11-hexadecadienyl acetate isomers, although larger females emit a slightly higher proportion of the Z,Z isomer. The synthetic pheromone used for disruption matches the average blend ratio (1:1 ZZ:ZE), assumed to represent the most abundant medium-sized females. We hypothesized that in pheromone-saturated environments, males would face difficulties locating medium-sized females whose pheromone resembles the synthetic blend. Consequently, we predicted reduced mating success for medium-sized females, potentially shifting body size distributions in the population. To test this, we compared the mating success of large, medium, and small females from a laboratory “naïve” population, never exposed to synthetic pheromone, and from a field population exposed to pheromone for many generations, with and without synthetic pheromone. We also measured pupal weights to assess body size differences. Results showed that medium-sized females experienced significantly reduced mating success in the presence of synthetic pheromone. Additionally, moths from mating disruption–treated fields were, on average, larger than those from the lab. Because larger females are more fecund, laying substantially more eggs than medium- or small-sized females, their increased mating success could partially counteract the suppression effect of the mating disruption strategy. Figures Figure 1 Figure 2 Figure 3 Introduction Male moths generally locate potential mates by detecting and following the pheromone plumes of females (Greenfield 1981 ; Cardé and Baker 1984 ). This pheromone-mediated communication is crucial for reproduction in most moth species (Cardé 2016 ). Leveraging this process, the mating disruption technique, widely used in agricultural pest control, deploys synthetic pheromone components in fields to interfere with males' ability to locate females (Cardé 1990 ; Cardé and Minks 1995 ; Howse et al. 1998 ). The reduced ability of males to detect receptive females may be explained by several mechanisms, including competition with synthetic pheromone sources, masking of female pheromone plumes, sensory adaptation (reducing the responsiveness of olfactory receptors), and habituation (a central nervous system process that reduces behavioral responses to female pheromones) (Cardé and Minks 1995 ; Saunders 1997; Miller and Gut 2015 ). The efficacy of this technique depends on various factors, including the formulation and release method of the synthetic pheromone and environmental conditions such as canopy structure and wind direction shifts (Cardé 1990 ; Cardé and Minks 1995 ; Lykouressis et al. 2005 ). In some moth species, both sexes release pheromones attractive to the opposite sex (Landolt and Heath 1990 ). In such species, disrupting only the male's orientation to the female may not fully prevent mating encounters (Cardé and Minks 1995 ). Additionally, the success of mating disruption can be reduced by migration of mated females into treated areas from external populations (Sexton and Il'ichev 2000; Saucke et al. 2014 ; Régnière et al. 2019 ). High pest densities can also diminish the effectiveness of this control method by increasing the likelihood of random encounters between males and females, amidst the synthetic pheromone plumes (Carpenter et al. 1982 ; Lance et al. 2016 ). The success of mating disruption techniques varies among species. Notable successes include the oriental fruit moth ( Grapholita molesta ), a key pest of stone and pome fruits (Rothschild 1975 ; Gentry et al. 1980 ; Vickers et al. 1985 ; Il’chev et al. 2004; Kong et al. 2014 ); the tomato pinworm ( Keiferia lycopersicella ), a major pest of tomatoes (Van Steenwyk et al. 1983; Wang et al. 1997; Schuster et al. 2000); the European grape moth ( Eupoecilia ambiguella ) and the grapevine moth ( Lobesia botrana ), both significant pests in European vineyards (Louis and Schirra 2001 ; Lioriatti et al. 2004 ; Benelli et al. 2023 ; Ricciardi et al. 2024 ); and the pink bollworm ( Pectinophora gossypiella ), a major pest of cotton worldwide (Gaston et al. 1977 ; Staten et al. 1987; Cardé 1990 ; Critchley et al. 1991 ; Collar et al. 2002; Lykouressis et al. 2005 ). The pink bollworm moth is a highly destructive cotton pest (Lukefahr and Martin 1963 ; Agarwal and Katiyar 1979 ; Ingram 1980 ; Parmar and Pate 2016; Bhute et al. 2023 ). Pink bollworm larvae primarily feed on developing cotton seeds, leading to premature boll opening or significant postharvest damage due to rotting (Parmar and Pate 2016; Sarwar 2017 ). The pink bollworm moth is challenging to manage with insecticides: Females predominantly lay eggs on the sutures or beneath the bracteoles at the base of bolls, and the larvae penetrate the bolls where they are protected from conventional insecticides (Lykouressis et al. 2005 ; Sreenivas et al. 2021). The pink bollworm’s sex pheromone was identified in 1973 as a 1:1 blend of cis/cis (Z,Z) and cis/trans (Z,E) isomers of 7,11-hexadecadienyl acetate (Hummel et al. 1973 ), though deviations of this ratio have been documented (Haynes et al. 1984 ; Gonzalez-Karlsson et al. 2021 ). Building on this discovery, studies in the late 1970s and early 1980s demonstrated promising results using 'Gossyplure', a synthetic formulation of this 1:1 blend, to disrupt the moths' mating (Gaston et al. 1977 ; Flint et al. 1978 ; Henneberry et al. 1981 ). Since then, mating disruption has been successfully used widely to control the pink bollworm moth in cotton fields (Brooks et al. 1979 ; Staten et al. 1987; Cardé and Minks 1995 ; Athanassiou et al. 2002 ; Collar et al. 2002; Lykouressis et al. 2005 ). However, recent field evidence indicates that the mating disruption faces significant challenges in controlling the pink bollworm (Niv 2000 ; Harari and Sharon 2022 ). Research has suggested that mating disruption alone may not adequately control pink bollworm populations, highlighting the necessity to integrate other pest management strategies (Cardé and Minks 1995 ; Niv 2000 ). Here, we propose an evolutionary factor that may influence the efficacy of this important, environmentally friendly control method. The synthetic pheromone used for mating disruption of the pink bollworm moth consists of a 1:1 blend of its two components, reflecting the average pheromone composition of pink bollworm in field populations. However, larger and younger females in good condition tend to produce a higher ratio of Z,Z to Z,E isomers in their pheromone glands, which is recognized and preferred by males (Gonzalez-Karlsson et al. 2021 ). We hypothesized that the synthetic pheromone, which likely mirrors the emission ratios of the most abundant females in the population (i.e., medium-sized females), would predominantly affect the mating success of these females. A lesser impact on larger and smaller females is expected since their pheromone blend deviates from the 1: 1 ratio. Over evolutionary timescales, the higher mating success of large and small females under continuous mating disruption could lead to shifts in body size distribution within treated populations. If mating disruption disproportionately reduces the mating success of medium-sized females, this could lead to a shift in the mating proportions among other size classes in the population, with important implications for overall fecundity and the long-term efficacy of the technique. Since larger females in naïve populations exhibit higher fecundity than smaller females (Gonzalez-Karlsson et al. 2021 ), we hypothesized that this pattern would persist in field populations exposed to mating disruption. Therefore, an increase in the proportion of large females in treated fields could potentially undermine the overall efficacy of the mating disruption technique. In this study, we tested whether the mating success of females, across different size classes (large, medium, or small), differs under conditions of pheromone excess compared to naïve populations. We also examined the body size distribution of moths collected from mating disruption environments versus lab-reared moths, and investigated female fecundity based on their body size categories. By addressing these questions, this research provides the first evolutionary insights into how sexual selection affects the efficacy of mating disruption. Our findings also lay the foundation for developing improved strategies aiming to optimize this environmentally friendly pest control method for sustainable agricultural practices. Methods Breeding and maintenance Two populations of pink bollworm moths were maintained in separate climate-controlled rooms at 25 ± 1°C, a 14:10 light-to-dark photoperiod and 60% relative humidity: (1) a naïve population collected before the implementation of mating disruption in cotton fields, never exposed to this method. These moths have been reared in the lab for approximately 40 years since their collection, and (2) a population established from larvae collected from cotton bolls in a field exposed to prolonged mating disruption. The latter group was reared in the laboratory for 10–15 generations before being used in our experiments, maintained continuously under a pheromone-saturated environment, using synthetic pheromone ropes (1:1 ratio of the two components, Shin-Etsu, Japan). Larvae from both groups were reared on an artificial diet (Stonefly Heliothis Diet, Ward’s Science), and adults were provided with a 10% sugar solution ad libitum. Males and females were separated during the larval stage into two cages. Females were grouped by size during the pupae stage into three size categories: large (0.0170–0.0220 g), medium (0.0120–0.0160 g), and small (0.0070–0.0110 g). Adult moths were separated daily into age-based (30 x 30 x 30 cm) screen cages. Experimental design Experiment 1: The effect of mating disruption on female mating success in relation to their body size. To test the hypothesis that mating disruption reduces the mating success of medium-sized females compared to naïve females, we conducted the following experiment: We placed 40 randomly sized males with 10 large, 10 medium, and 10 small females in a net cage. The insects were allowed to interact for 40 minutes during their mating window. Once a pair mated, they were removed, and the female’s size category (large, medium, or small) was identified using a pre-established visual scale. The scale was based on size of adult females that emerged from selected pupae of the different size groups (large, medium and small). This scale allowed us to reliably determine each female's category at the time of pair removal. The mated female was then replaced with another female of the same size category, while a new male was randomly selected to replace the removed male. This ensured a constant representation of all female size categories throughout the experiment. This experiment included the following three treatments: Naïve moths that have never been exposed to mating disruption, without synthetic pheromone during the experiment ( n = 17 trials). Prolonged pheromone-exposed moths in the presence of synthetic pheromone during the experiment ( n = 15 trials). Prolonged pheromone-exposed moths in the absence of synthetic pheromone during the experiment ( n = 15 trials). Note that naïve moths were not tested under mating disruption conditions. Previous studies showed that first exposure of naïve moths to synthetic pheromone causes a marked reduction in activity (Waner et al. 2025), making behavioral comparisons difficult and uninformative for the purpose of this study. We evaluated the proportion of medium-sized mated females across the three treatments by comparing the proportions of large versus medium-sized females and medium versus small-sized females. Statistical analysis L − M (Large minus Medium) represents the difference in proportions between large- and medium-sized mated females. Across all three treatments, the distribution of L − M was normal; therefore, t-tests were used to compare L − M between treatments. Similarly, M − S (Medium minus Small) represents the difference in proportions between medium- and small-sized mated females. Since the distribution of M − S was not normal for all treatments, comparisons of M − S were conducted using the non-parametric Mann-Whitney U -test. Note that we were only interested in comparing medium-sized females to large- and small-sized females (but not large to small). Thus, we restricted our analysis to L − M and M − S to maximize statistical power. Comparative Analysis 1: Comparing the size distribution of moths from naïve and pheromone-exposed populations. To evaluate the differences in the size distribution between naïve moths and moths with prolonged exposure to mating disruption, we measured the weights of pupae randomly collected from the following three groups: Naïve Population : Males ( n = 107) and females ( n = 97) from a population that has never been exposed to mating disruption. Field-Collected Population : Males ( n = 136) and females ( n = 143) collected as final-instar larvae from a cotton field treated with mating disruption. Laboratory-Reared Field Population : Males ( n = 172) and females ( n = 183) from the same field-collected population as in group 2, reared for two generations under laboratory conditions similar to the naïve population but with the presence of synthetic pheromone (Shin-Etsu, Japan) in the rearing room. This approach controlled for the potential effect of laboratory diet on moth body size. statistical analysis The weights of the 423 female and 415 male pupae were normally distributed. To assess potential weight differences based on origin, we conducted a one-way Welch's ANOVA for both female and male pupae, as this method is robust to unequal variances. In order to evaluate the overall significance of the weight differences between female and male pupae across all treatment groups, we employed Winer’s method of adding t -values (see Rosenthal, 1978). Comparative Analysis 2: The impact of female size on fecundity in mating disruption-exposed populations. Previous research has shown that larger naïve moths are more fecund than smaller naïve moths. To test whether female size affects fecundity in mating disruption-exposed females, we counted the number of eggs laid by groups of 10 females classified as large ( n = 6 groups), medium ( n = 6 groups), or small ( n = 5 groups). Each group was kept in a mating container with 15 randomly sized males. The excess of males aimed to ensure mating for all females. Egg-laying substrates were replaced twice weekly, and eggs were counted. Following the death of the females, dissections were performed to confirm the mating status of the females by the presence of at least one spermatophore. Statistical analysis The number of eggs did not have a normal distribution (Kolmogorov-Smirnov test: p = 0.037). Thus, a square-root transformation was applied for the one-way ANOVA analysis. Results Experiment 1: The effect of mating disruption on female mating success in relation to their body size. Proportion of mated females: In the naïve population that was never exposed to synthetic pheromone, 47.41% of medium-sized females mated, compared to 39.26% of large females and 13.33% of small females. In contrast, in prolonged pheromone-exposed moths in the presence of synthetic pheromone, only 25.93% of medium-sized females mated, while 55.56% of large females and 18.52% of small females mated. For moths that were previously exposed to prolonged mating disruption but were not exposed to synthetic pheromone during the experiment, 43.09% of medium-sized females, 50.41% of large females, and 6.50% of small females successfully mated. There was no significant difference in L − M between the naïve population and the moths long exposed to mating disruption in the absence of synthetic pheromone ( t 30 = 0.320, p = 0.751), with both groups showing similar mating percentages for large and medium-sized females. However, significant differences in L − M were found between the naïve population and the prolonged pheromone-exposed moths in the presence of the synthetic pheromone ( t 30 = 2.838, p = 0.008), and between the prolonged pheromone-exposed moths in the presence vs. the absence of synthetic pheromone during the experiment ( t 28 = −2.703, p = 0.012), due to a lower proportion of mated medium-sized females in the presence of synthetic pheromone (Fig. 1 ). There was no significant difference in M − S between the naïve population and the prolonged pheromone-exposed moths in the absence of synthetic pheromone ( p = 0.830). However, significant differences were observed between the naïve population vs. the prolonged pheromone-exposed moths with synthetic pheromone ( p < 0.001), as well as between the prolonged pheromone-exposed moths in the presence vs. the absence of synthetic pheromone during the experiment ( p < 0.001), again due to a lower proportion of medium-sized females mating in the presence of synthetic pheromone (Fig. 1 ). Figure 1 . Percentage (average ± se) of mated females in the three treatments: A. Difference between large and medium sized ( L–M ) females. B. Difference between medium and small sized (M–S ) females. Comparative Analysis 1: Comparing the size distribution of moths in naïve and pheromone-exposed populations. Pupae, males, and females originating from a cotton field under mating disruption and subsequently reared in the laboratory for two generations in the presence of synthetic pheromone were significantly heavier than naïve moths (both males and females) that had never been exposed to synthetic pheromone (Table 1 , Fig. 2 ). Across all groups, females were consistently larger than males (p < 0.001; Fig. 2 ). Table 1 Multiple comparisons of weights by origin (Games-Howell method) Multiple comparisons Females Males Naïve vs. Field-Collected p < 0.001 p < 0.001 Naïve vs. field, reared in the Laboratory p < 0.001 p < 0.001 Field-Collected vs. field, reared in the Laboratory p = 0.905 p = 0.950 Figure 2 : Mean (± SE) pupal weights of female and male moths from three population origins: naïve population, field-collected population (F0), and field-collected larvae reared in the laboratory for two generations under mating disruption conditions (F2). Orange bars represent females; blue bars represent males. Different Latin letters indicate statistically significant differences among female groups, while different Greek letters indicate significant differences among male groups. Comparative Analysis 2: The impact of female size on fecundity in mating disruption-exposed populations. Larger females laid a significantly higher number of eggs than medium-size and small females. One-way ANOVA: p < 0.001. Multiple comparisons: Small vs. Medium p = 0.087, Small vs. Large p < 0.001, Medium vs. Large p = 0.039. (Fig. 3 ). Discussion Mating disruption is an environmentally friendly tactic used to control numerous moth pest species, including the pink bollworm moth in cotton fields (Cardé and Minks 1995 ; Staten et al. 1997 ). This species-specific approach minimizes harm to non-target organisms within the treated area. Instead of lethally targeting pests, it interferes with mating, thereby reducing reproductive success, lowering population levels, and ultimately decreasing crop damage (Mevada et al. 2023 ). While this technique has been largely effective against the pink bollworm, there remains room for improvement in ensuring its long-term efficacy. Analyses of the proportions of mated females across size categories revealed significant differences between pheromone-exposed moths, tested in the presence of synthetic pheromone, and naïve laboratory-reared moths. Similarly, significant size-related differences were also observed within the pheromone-exposed group when comparing trials conducted with and without synthetic pheromone. In contrast, no such differences were detected between laboratory-reared moths and pheromone-exposed moths when both were tested in the absence of synthetic pheromone. The proportion of small-mated females was consistently the lowest across all three experimental conditions. In the naïve population, which had never been exposed to mating disruption, medium-sized females mated at rates comparable to, or slightly higher than large females. Given that larger females generally have higher fecundity and are typically preferred by males (Gonzalez-Karlsson et al. 2021 ), this suggests that mating success is shaped not only by male preference for high-quality mates but also by female selectivity. Larger, higher-quality pink bollworm females tend to be more selective, while smaller females are less preferred by males (Waner Rips, submitted). As a result, medium-sized females, though not the highest-quality mates, may achieve relatively high mating success by being less selective than large females and more attractive than small ones. Interestingly, the proportion of mated medium-sized females dropped sharply under pheromone-treated conditions, influencing the mating proportions of both large and small females. These shifts are reflected in the differences in the mating rate between large and medium females (L − M), and between medium and small females (M − S) (Fig. 1 ). These findings suggest that the standardized 1:1 ratio of pheromone components used in mating disruption exerts size-dependent effects, disproportionately reducing the mating success of medium-sized females. A previous study (Gonzalez-Karlsson et al. 2021 ) showed that large females produce a higher proportion of the Z,Z isomer, while small females produce more of the Z,E isomer in their sex pheromone blends. Both deviate from the 1:1 (Z,Z):(Z,E) ratio used in the commercial mating disruption formulation (Shin-Etsu, Japan). Although the pheromone blends of females in our study were not directly analyzed, large and small females likely retained their respective isomer biases. Medium-sized females, being the most prevalent in the population, likely emitted blends closer to the synthetic 1:1 ratio, rendering them more susceptible to pheromone masking. This may explain their reduced mating success under disrupted conditions, while the less common blends of large and small females were less affected. Another notable finding of this study is that moths collected from pheromone-treated cotton fields were significantly larger than laboratory-reared moths. This size difference persisted even after two generations of rearing under identical laboratory conditions. Both sexual selection and natural selection (including environmental changes) generally influence the body size of insects; Larger female insects typically achieve greater reproductive success (Wickman and Karlsson 1989 ), thus being favored by sexual selection processes. This can drive a shift toward larger female body size, particularly in species where larger females are strongly preferred by males (Andersson and Iwasa 1996 ). Similarly, sexual selection tends to favor larger males, as they often gain a competitive advantage in securing mates, thereby achieving more copulations (Alcock 1994 ; Serrano-Meneses et al. 2007 ). The reproductive advantage of large males and females is well-documented across taxa (Olsson et al. 2002 ; Byrne and Rice 2006 ). At the same time, natural selection often counterbalances this trend by favoring intermediate phenotypes, such as average body size, over extreme ones (Blanckenhorn 2000 ). Selection for intermediate body size, a form of stabilizing selection, occurs when extreme sizes come with fitness disadvantages. Smaller females often have reduced fecundity and energy reserves, while larger females may experience increased energy demands, higher developmental costs, greater predation risk (Kullberg et al. 1996 ; Neems et al. 1998 ; Blanckenhorn 2000 ; Covas et al. 2002 ), or an overall higher mortality rate (Xu and Wang 2013 ). Environmental changes can shift body size distributions in insects, as demonstrated for climate-driven trends in various species (Brose et al. 2012 ). In our study, we propose that the synthetic pheromone may weaken stabilizing selection in disrupted populations by disproportionately reducing the mating success of medium-sized females. Since males rely on pheromonal cues to locate mates, and medium-sized females are more likely to emit blends resembling the synthetic 1:1 ratio used in disruption, their signals may be more effectively masked. As a result, large females, whose pheromone blends deviate from the synthetic standard, may achieve higher mating rates, potentially shifting body size distributions in the population. Furthermore, the masking of medium-sized females may increase mating opportunities for smaller, less-preferred females, as males lower their selectivity when mate location becomes more challenging. Consistent with this, we observed a slight increase in the mating success of small females under pheromone-saturated conditions. However, because small females are generally less preferred, this shift likely contributes less to selection pressures than the increased mating of large females. Environmental factors such as diet or population density may also affect body size. Our naïve population had been reared in the laboratory for ~ 40 years, which could have influenced female size, although previous studies suggest long-term lab rearing has little effect on lepidopteran body size (Hoffmann and Ross 2018 ). Still, we cannot exclude the possibility that it also contributed to the smaller sizes observed in naïve moths compared with field-collected moths. To specifically address the potential influence of laboratory diet, one of the main factors that could affect lab moths’ size, we reared a subset of the field-collected population in the laboratory for two additional generations under controlled conditions, with synthetic pheromones present. The pupae from this second generation were also significantly larger than those from the naïve population, and their size did not differ from that of pupae collected directly from treated fields. Insect body size is influenced by genetic and environmental factors and plays a critical role in determining fitness (Nijhout 2003 ; Edgar 2006 ; Beukeboom 2018 ; Nijhout et al. 2003). Although specific data disentangling the genetic and environmental influences on body size in the pink bollworm is unavailable, both factors are expected to play a role. If this pattern persists across generations, it could have contributed to an increase in the average size of moths in treated areas. Female size in insects is often a reliable indicator of fecundity (Honěk 1993 ). Large female pink bollworm moths that have never been exposed to mating disruption are more fecund than smaller moths (Gonzalez-Karlsson et al. 2021 ). We re-tested this relationship in moths from a field treated with mating disruption for many generations. Similarly, we found that larger females were significantly more fecund than medium-sized and small females, as measured by the total number of eggs laid over their lifetimes. A higher proportion of larger females, coupled with their higher fecundity, in areas subjected to mating disruption, raises questions about the long-term efficacy of this method. If larger females consistently contribute more offspring, this could influence population structure dynamics and potentially reduce the effectiveness of mating disruption over time. To address this challenge, exploring and applying alternative ratios of sex pheromone isomers could enhance the efficacy of this promising and environmentally friendly approach. Further research into the interplay between female size, fecundity, and pheromone communication under mating disruption conditions may provide valuable insights, contributing to the refinement of this method for sustainable pest management. Declarations Ethical Note The pink bollworm moth ( Pectinophora gossypiella ) is a well-known agricultural pest. Consequently, research involving this species typically does not require specific ethical permits. Nonetheless, all experimental procedures were conducted in adherence to ethical standards. The insects were reared in climate-controlled rooms maintained at a constant temperature of 25°C and were consistently provided with appropriate food corresponding to their developmental stages. Throughout the study, the moths were handled gently and with care. After mating trials, they were released. At no point were the insects harmed or subjected to detrimental conditions. Author Contribution SWR conceived the research questions and designed the experiments with the help of AH and MM. SWR conducted the experiments, analyzed the data, and wrote the manuscript. AH and MM assisted in experiment design, conducted the experiments, and reviewed the manuscript. UM contributed to the statistical analysis and reviewed the manuscript. OK took part in conceiving the research questions and revising the manuscript. All authors read and approved the final version of the manuscript. Acknowledgement We thank Arnon Lotem and Victoria Soroker for their valuable advice and insightful discussions that greatly improved this study. We are also deeply grateful to Prof. Ariel Chipman for his continuous support and thoughtful advice whenever needed. Our thanks extend to Eyal Halon, Michael Davidivitz, Nikolay Meltser, and Aya Rafael Cohen for their dedicated technical assistance and field support. This research was partially supported by the Israel Science Foundation (Grant No. 1826/20), the British Friends of the Hebrew University, and the Yael Pitoun Scholarship. Data Availability The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request. References Agarwal RA, Katiyar KN (1979) An estimate of losses of seed kapas and seed due to bollworms on cotton in India. Indian J Entomol 41:143–148 Alcock J (1994) Body size and its effect on male-male competition in Hylaeus alcyoneus (Hymenoptera: Colletidae). J Insect Behav 8:149–159 https://doi.org/10.1007/BF01988901 Andersson M, Iwasa Y (1996) Sexual selection. Trends Ecol Evol 11:53–58 https://doi.org/10.1016/0169-5347(96)81042-1 Athanassiou CG, Buchelos CT, Kavallieratos NG, Barbetaki AE (2002) Evaluation of the mating disruption method for the control of the pink bollworm Pectinophora gossypiella (Saund.) (Lepidoptera: Gelechiidae) and comparison of this method with insecticidal treatments. IOBC WPRS Bull 25 Benelli G, Lucchi A, Anfora G, Bagnoli B, Botton M, Campos-Herrera R et al. (2023) European grapevine moth, Lobesia botrana . Part II: Prevention and management. Entomologia Generalis 43(2):281–304 https://doi.org/10.1127/entomologia/2023/1947 Beukeboom LW (2018) Size matters in insects: An introduction. Entomologia Experimentalis et Applicata 166(1):2–3 https://doi.org/10.1111/eea.12646 Bhute NK, Patil CS, Deshmukh KV, Wagh RS, Medhe NK (2023) Pink bollworm Pectinophora gossypiella (Saunders), a destructive pest of cotton: a review. Pharma Innovation J 12(3):2036–2042 Blanckenhorn WU (2000) The evolution of body size: what keeps organisms small? Quart Rev Biol 75(4):385–407 https://doi.org/10.1086/393620 Brooks TW, Doane CC, Staten RT (1979) Experience with the first commercial pheromone communication disruptor for suppression of an agricultural insect pest. In: Ritter FJ (ed) Chemical Ecology: Odour Communication in Animals . Elsevier/North-Holland Biomedical Press, Amsterdam, pp 375–388 Brose U, Dunne JA, Montoya JM, Petchey OL, Schneider FD, Jacob U (2012) Climate change in size-structured ecosystems. Phil Trans R Soc B 367:2903–2912 https://doi.org/10.1098/rstb.2012.0232 Byrne PG, Rice WR (2006) Evidence for adaptive male mate choice in the fruit fly Drosophila melanogaster . Proc R Soc B 273:917–922 https://doi.org/10.1098/rspb.2005.3372 Cardé RT, Baker TC (1984) Sexual communication with pheromones. In: Bell WJ, Cardé RT (eds) Chemical Ecology of Insects . Springer US, Boston MA, pp 355–383 Cardé RT (1990) Principles of mating disruption. In: Ridgway RL, Silverstein RM, Inscoe MN (eds) Behavior-Modifying Chemicals for Pest Management: Applications of Pheromones and Other Attractants . Marcel Dekker, New York, pp 47–71 Cardé RT, Minks AK (1995) Control of moth pests by mating disruption: successes and constraints. Annu Rev Entomol 40:559–585 https://doi.org/10.1146/annurev.en.40.010195.003015 Cardé RT (2016) Moth navigation along pheromone plumes. In: Allison JD & Cardé RT (eds) Pheromone Communication in Moths: Evolution, Behavior, and Application . University of California Press, Berkeley, pp 173–190 https://doi.org/10.1525/9780520964433-012 Carpenter JE, Sparks AN, Gueldner RC (1982) Effects of moth population density and pheromone concentration on mating disruption of the corn earworm in large screened cages. J Econ Entomol 75:333–336 https://doi.org/10.1093/jee/75.2.333 Collar Urquijo JL, Gálvez Manzano V (2002) Mating disruption strategies for the control of pink bollworm ( Pectinophora gossypiella Saunders) in cotton fields. Boletín Sanid Veg Plagas 28:193–198 Covas R, Brown CR, Anderson MD, Brown MB (2002) Stabilizing selection on body mass in the sociable weaver Philetairus socius . Proc R Soc B 269:1905–1909 https://doi.org/10.1098/rspb.2002.2106 Critchley BR, Chamberlain DJ, Campion DG, Attique MR, Ali M, Ghaffar A (1991) Integrated use of pink bollworm pheromone formulations and selected conventional insecticides for the control of the cotton pest complex in Pakistan. Bull Entomol Res 81:371–378 Edgar BA (2006) How flies get their size: genetics meets physiology. Nat Rev Genet 7:907–916 https://doi.org/10.1038/nrg1989 Flint HM, Butler LI, McDonough LM, Smith RL, Forey DE (1978) Pink bollworm: response to various emission rates of gossyplure in the field. Environ Entomol 7:57–61 https://doi.org/10.1093/ee/7.1.57 Gaston LK, Kaae RS, Shorey HH, Sellers D (1977) Controlling the pink bollworm by disrupting sex pheromone communication between adult moths. Science 196:904–905 https://doi.org/10.1126/science.870967 Gentry CR, Bierl-Leonhardt BA, Blythe JL, Plimmer JR (1980) Air permeation tests with orfralure for reduction in trap catch of oriental fruit moths. J Chem Ecol 6:185–192 Gonzalez-Karlsson A, Golov Y, Steinitz H, Moncaz A, Halon E, Horowitz AR, Goldenberg I, Gurka R, Liberzon A, Soroker V, Jurenka R (2021) Males perceive honest information from female-released sex pheromone in a moth. Behav Ecol 32:1127–1137 https://doi.org/10.1093/beheco/arab073 Greenfield MD (1981) Moth sex pheromones: an evolutionary perspective. Fla Entomol 64:4–17 https://doi.org/10.2307/3494597 Harari A, Sharon R (2022) The contemporary and prospective risks of resistance to the mating disruption method in moths. Entomol Gen 42(2):275–288 https://doi.org/10.1127/entomologia/2021/1275 Haynes KF, Gaston LK, Pope MM, Baker TC (1984) Potential for evolution of resistance to pheromones: interindividual and interpopulational variation in the chemical communication system of the pink bollworm moth. J Chem Ecol 10:1551–1565 https://doi.org/10.1007/BF00988424 Honěk A (1993) Intraspecific variation in body size and fecundity in insects: a general relationship. Oikos 66(3):483–492 https://doi.org/10.2307/3544943 Henneberry TJ, Gillespie JM, Bariola LA, Flint HM, Lingren PD, Kydonieus AF (1981) Gossyplure in laminated plastic formulations for mating disruption and pink bollworm control. J Econ Entomol 74(4):376–381 https://doi.org/10.1093/jee/74.4.376 Hoffmann AA, Ross PA (2018) Rates and patterns of laboratory adaptation in (mostly) insects. J Econ Entomol 111(2):501–509 https://doi.org/10.1093/jee/toy024 Howse PE, Stevens IDR, Jones OT (1998) Mating disruption. In: Howse PE, Stevens IDR, Jones OT (eds) Insect Pheromones and Their Use in Pest Management. Springer, Dordrecht, pp 314–344 https://doi.org/10.1007/978-94-011-5344-7_12 Hummel HE, Gaston LK, Shorey HH, Kaae RS, Byrne KJ, Silverstein RM (1973) Clarification of the chemical status of the pink bollworm sex pheromone. Science 181(4102):873–875 https://doi.org/10.1126/science.181.4102.873 Ingram WR (1980) Studies of the pink bollworm, Pectinophora gossypiella, on sea island cotton in Barbados. Int J Pest Manag 26(2):118–137 https://www.tandfonline.com/doi/abs/10.1080/09670878009414384 Kong WN, Li J, Fan RJ, Li SC, Ma RY (2014) Sex-pheromone-mediated mating disruption technology for the oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae): overview and prospects. Psyche A J Entomol 2014:253924 https://doi.org/10.1155/2014/253924 Kullberg C, Fransson T, Jakobsson S (1996) Impaired predator evasion in fat blackcaps ( Sylvia atricapilla ). Proc R Soc B 263(1377):1671–1675 https://doi.org/10.1098/rspb.1996.0244 Landolt PJ, Heath RR (1990) Sexual role reversal in mate-finding strategies of the cabbage looper moth. Science 249(4972):1026–1028 https://doi.org/10.1126/science.249.4972.1026 Lance DR, Leonard DS, Mastro VC, Walters ML (2016) Mating disruption as a suppression tactic in programs targeting regulated lepidopteran pests in US. J Chem Ecol 42(7):590–605 https://doi.org/10.1007/s10886-016-0732-9 Louis F, Schirra KJ (2001) Mating disruption of Lobesia botrana (Lepidoptera: Tortricidae) in vineyards with very high population densities. IOBC WPRS Bull 24(2):75–80 Ioriatti CL, Bagnoli BR, Lucchi AN, Veronelli VI (2004) Vine moths control by mating disruption in Italy: results and future prospects. Redia 87:117–128 Il’ichev AL, Williams DG, Milner AD (2004) Mating disruption barriers in pome fruit for improved control of oriental fruit moth Grapholita molesta (Busck) in stone fruit under mating disruption. J Appl Entomol 128(2):126–132 https://doi.org/10.1111/j.1439-0418.2004.00822.x Lukefahr MJ, Martin DF (1963) Evaluation of damage to lint and seed of cotton caused by the pink bollworm. J Econ Entomol 56(5):710–713 Lykouressis D, Perdikis D, Samartzis D, Fantinou A, Toutouzas S (2005) Management of the pink bollworm Pectinophora gossypiella (Saunders) by mating disruption in cotton fields. Crop Prot 24(2):177–183 https://doi.org/10.1016/j.cropro.2004.07.007 Mevada RR, Sisodiya DB, Parmar RG, Prajapati DR (2023) Mating disruption: an ecological step towards sustainable pest management. J Eco-Friendly Agric 18(1):144–150 https://doi.org/10.5958/2582-2683.2023.00025.4 Miller JR, Gut LJ (2015) Mating disruption for the 21st century: matching technology with mechanism. Environ Entomol 44(3):427–453 https://doi.org/10.1093/ee/nvv052 Neems RM, Lazarus J, McAdllan AJ (1998) Lifetime reproductive success in a swarming midge: trade-offs and stabilizing selection for male body size. Behav Ecol 9(3):279–286 https://doi.org/10.1093/beheco/9.3.279 Nijhout HF (2003) The control of body size in insects. Dev Biol 261(1):1–9 https://doi.org/10.1016/S0012-1606(03)00276-8 Nijhout HF, Riddiford LM, Mirth CK, Shingleton A, Suzuki Y, Callier V (2014) The developmental control of size in insects. WIREs Dev Biol 3(1):113–134 https://doi.org/10.1002/wdev.124 Niv A (2000) Use of pheromones for pink bollworm ( Pectinophora gossypiella , Saunders) mating disruption in Israel. In: Proceedings of the World Cotton Research Conference, Vol 2, pp 6–12 Olsson M, Shine R, Wapstra E, Ujvari B, Madsen T (2002) Sexual dimorphism in lizard body shape: the roles of sexual selection and fecundity selection. Evolution 56(7):1538–1542 https://doi.org/10.1111/j.0014-3820.2002.tb01464.x Parmar VR, Patel CC (2016) Pink bollworm: a notorious pest of cotton: a review. AGRES Int e-J 5(2):88–97 Régnière J, Delisle J, Dupont A, Trudel R (2019) The impact of moth migration on apparent fecundity overwhelms mating disruption as a method to manage spruce budworm populations. Forests 10(9):775 https://doi.org/10.3390/f10090775 Ricciardi R, Benelli G, Di Giovanni F, Lucchi A (2024) The European grape berry moth, Eupoecilia ambiguella (Lepidoptera: Tortricidae): Current knowledge and management challenges. Crop Prot p 106641 https://doi.org/10.1016/j.cropro.2024.106641 Rothschild GHL (1975) Control of oriental fruit moth ( Cydia molesta (Busck)) (Lepidoptera: Tortricidae) with synthetic female pheromone. Bull Entomol Res 65(3):473–490 https://doi.org/10.1017/S0007485300006143 Saucke H, Balasus A, Finckh MR, Formowitz B, Schmidt R, Kratt A (2014) Mating disruption of pea moth ( Cydia nigricana ) in organic peas ( Pisum sativum ). Entomol Exp Appl 150(3):199–207 https://doi.org/10.1111/eea.12153 Sarwar M (2017) Biological parameters of pink bollworm Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae): a looming threat for cotton and its eradication opportunity. Int J Res Agric For 4(7):25–36 Serrano-Meneses MA, Córdoba-Aguilar A, Méndez V, Layen SJ, Székely T (2007) Sexual size dimorphism in the American rubyspot: male body size predicts male competition and mating success. Anim Behav 73(6):987–997 https://doi.org/10.1016/j.anbehav.2006.08.01 Staten RT, El-Lissy O, Antilla L (1997) Successful area-wide program to control pink bollworm by mating disruption. In: Cardé RT, Minks AK (eds) Insect Pheromone Research: New Directions . Springer US, Boston MA, pp 383–396 Vickers RA, Rothschild GHL, Jones EL (1985) Control of the oriental fruit moth, Cydia molesta (Busck) (Lepidoptera: Tortricidae), at a district level by mating disruption with synthetic female pheromone. Bull Entomol Res 75(4):625–634 https://doi.org/10.1017/S0007485300015881 Wickman PO, Karlsson B (1989) Abdomen size, body size and the reproductive effort of insects. Oikos 56(2):209–214 https://doi.org/10.2307/3565338 Xu J, Wang Q (2013) Trade-off between adult body size and juvenile survival: an experimental test of parental effects in the Mediterranean flour moth. Aust J Entomol 52:403–406 https://doi.org/10.1111/aen.12038 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7842351","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":550041068,"identity":"35283669-c2ce-4be9-adb9-6d65b4f4b623","order_by":0,"name":"Shevy Waner Rips","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAxElEQVRIiWNgGAWjYDACCQglByIOPCBFizFYSwIpWhIbQCRRWuRnNx+T5m07nD4/7PBDoC12croNBLQY3DmWBtKSu/F2mgFQS7Kx2QFCWiRyjI15t93O3Tg7AaTlQOI2QlrkZ+R/BmlJN5yd/oE4LQw3chgfA7UkyEvnEGmLwY00w4dz//033CCdU3AgwYAIv8jPSH5w4M2ZNHn52embP3yosJMjqAUEmHhA1oFVGhChHAQYf4CsayBS9SgYBaNgFIw8AACp6Ei7dWoY/AAAAABJRU5ErkJggg==","orcid":"","institution":"Hebrew University of Jerusalem","correspondingAuthor":true,"prefix":"","firstName":"Shevy","middleName":"Waner","lastName":"Rips","suffix":""},{"id":550041069,"identity":"aaef4b51-edcd-4c6c-8b86-9e397df43def","order_by":1,"name":"Michal Motro","email":"","orcid":"","institution":"David Yellin College of Education","correspondingAuthor":false,"prefix":"","firstName":"Michal","middleName":"","lastName":"Motro","suffix":""},{"id":550041070,"identity":"2deae937-752f-4ca0-809d-44e0aee5e11a","order_by":2,"name":"Uzi Motro","email":"","orcid":"","institution":"Hebrew University of Jerusalem","correspondingAuthor":false,"prefix":"","firstName":"Uzi","middleName":"","lastName":"Motro","suffix":""},{"id":550041071,"identity":"d44b23af-78ef-4170-a83f-abbad7777f98","order_by":3,"name":"Oren Kolodny","email":"","orcid":"","institution":"Hebrew University of Jerusalem","correspondingAuthor":false,"prefix":"","firstName":"Oren","middleName":"","lastName":"Kolodny","suffix":""},{"id":550041072,"identity":"6f0ac378-26ba-456f-b137-f4cba6df40b7","order_by":4,"name":"Ally Harari","email":"","orcid":"","institution":"Volcani Institute","correspondingAuthor":false,"prefix":"","firstName":"Ally","middleName":"","lastName":"Harari","suffix":""}],"badges":[],"createdAt":"2025-10-12 17:53:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7842351/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7842351/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":97137402,"identity":"67a88f59-c91b-4f63-95bd-9b14dc5e478d","added_by":"auto","created_at":"2025-12-01 09:57:42","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":488567,"visible":true,"origin":"","legend":"","description":"","filename":"article324.8.25.docx","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/71f74ca8da2d664942e931ba.docx"},{"id":97136692,"identity":"b3216821-23d4-494c-bee6-00e5606113c9","added_by":"auto","created_at":"2025-12-01 09:56:53","extension":"json","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":7692,"visible":true,"origin":"","legend":"","description":"","filename":"2ab3df38ef8f4cfeafec5ad98f509487.json","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/30b5da48cfd579bd400c598d.json"},{"id":96944499,"identity":"59baa392-dd73-4f64-9b04-4a7cec7359bf","added_by":"auto","created_at":"2025-11-27 19:29:33","extension":"xml","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":128439,"visible":true,"origin":"","legend":"","description":"","filename":"2ab3df38ef8f4cfeafec5ad98f5094871enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/ebe22693d9a20eb6ecd3c323.xml"},{"id":96944496,"identity":"a27b46d5-fb13-43d9-92ae-d3896b2e5258","added_by":"auto","created_at":"2025-11-27 19:29:33","extension":"jpeg","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":29854,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/e311bee3d6d90e06c9c05208.jpeg"},{"id":97135832,"identity":"84c4af69-763b-4c8e-9e1c-97e9552a3f1c","added_by":"auto","created_at":"2025-12-01 09:53:57","extension":"jpeg","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":268678,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/f2ae0e19281723eef52da149.jpeg"},{"id":96944515,"identity":"728908ef-0a03-4abe-80c6-20ca4f745946","added_by":"auto","created_at":"2025-11-27 19:29:34","extension":"jpeg","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1074,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/8714ef9a0be6d052e8dff07d.jpeg"},{"id":96944514,"identity":"136bc524-93eb-45bd-8f07-67342080b44e","added_by":"auto","created_at":"2025-11-27 19:29:34","extension":"jpeg","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":177330,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/0675ea9217ddfbb0d11b043f.jpeg"},{"id":97136493,"identity":"daecb827-0cf1-4454-b176-028a3ea1e241","added_by":"auto","created_at":"2025-12-01 09:56:39","extension":"jpeg","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":116484,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/6eb3ed264255cc9bc10437cb.jpeg"},{"id":96944503,"identity":"e8f3f672-b786-433b-bc70-aac6d9ca7c22","added_by":"auto","created_at":"2025-11-27 19:29:33","extension":"jpeg","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":30117,"visible":true,"origin":"","legend":"","description":"","filename":"groupimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/b01096b7564abae0c163d360.jpeg"},{"id":96944512,"identity":"04ac0ffc-7b2e-4c04-834f-30d2c0094707","added_by":"auto","created_at":"2025-11-27 19:29:34","extension":"png","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":3878,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/4515a2c6cc54b4ecae2380e1.png"},{"id":97135844,"identity":"d95b4f95-2a84-4806-a871-0dabbead324e","added_by":"auto","created_at":"2025-12-01 09:54:00","extension":"png","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":58249,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/4a90aafc26db87be62448b41.png"},{"id":96944508,"identity":"d7f2d8a3-ec03-4096-acf1-502685077a87","added_by":"auto","created_at":"2025-11-27 19:29:33","extension":"png","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":935,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/9ddbc2bffdeaed0aa83acb0f.png"},{"id":96944504,"identity":"b5d3d078-c21f-490b-9fc1-11d17e14a2d2","added_by":"auto","created_at":"2025-11-27 19:29:33","extension":"png","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":29980,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/11b9bb2f7325bb745d0f830c.png"},{"id":97136949,"identity":"281817d2-0548-4431-9545-807e09b29168","added_by":"auto","created_at":"2025-12-01 09:57:12","extension":"png","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":22549,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/335814370fdee04275923483.png"},{"id":96944513,"identity":"21ef403a-9298-4817-8d0b-ab127f1da89a","added_by":"auto","created_at":"2025-11-27 19:29:34","extension":"png","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":10953,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinegroupimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/6274cb327201ff3459e30705.png"},{"id":96944511,"identity":"6218d34c-f126-4ab5-9478-61866b0fdf2b","added_by":"auto","created_at":"2025-11-27 19:29:34","extension":"xml","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":125533,"visible":true,"origin":"","legend":"","description":"","filename":"2ab3df38ef8f4cfeafec5ad98f5094871structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/3b0fa5e63f588ebf7cdec134.xml"},{"id":96944509,"identity":"6671f462-9d93-4193-a386-b8143bae2a21","added_by":"auto","created_at":"2025-11-27 19:29:33","extension":"html","order_by":16,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":136474,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/96869a3cff7e2157865d5cf5.html"},{"id":96944495,"identity":"60e77275-ab8e-4dd8-b2e4-8094101df837","added_by":"auto","created_at":"2025-11-27 19:29:33","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":121431,"visible":true,"origin":"","legend":"\u003cp\u003ePercentage (average ± se) of mated females in the three treatments: \u003cstrong\u003eA.\u003c/strong\u003e Difference between large and medium sized (\u003cem\u003eL–M\u003c/em\u003e) females. \u003cstrong\u003eB.\u003c/strong\u003e Difference between medium and small sized \u003cem\u003e(M–S\u003c/em\u003e) females.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/6fd32da9b3e75038ba239c63.png"},{"id":97136576,"identity":"d8dde0c4-b113-4942-af3b-2f69076c3e46","added_by":"auto","created_at":"2025-12-01 09:56:47","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":24977,"visible":true,"origin":"","legend":"\u003cp\u003eMean (± SE) pupal weights of female and male moths from three population origins: naïve population, field-collected population (F0), and field-collected larvae reared in the laboratory for two generations under mating disruption conditions (F2). Orange bars represent females; blue bars represent males. Different Latin letters indicate statistically significant differences among female groups, while different Greek letters indicate significant differences among male groups.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/af654a938b1254becefcabb7.png"},{"id":96944497,"identity":"4711a2e1-ea14-4bde-b2e2-44885b450558","added_by":"auto","created_at":"2025-11-27 19:29:33","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":14184,"visible":true,"origin":"","legend":"\u003cp\u003eThe average (+se) number of eggs laid by groups of 10 females according to size group: small females (0.0070-0.0110 g), medium sized females (0.0120-0.0160 g) and large females (0.0170-0.0220 g).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/bb1aef2664e6a943a5e265f5.png"},{"id":101655582,"identity":"99ccc01b-b223-4490-ae80-44cbacfb86bb","added_by":"auto","created_at":"2026-02-02 09:57:34","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":963254,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7842351/v1/b76906cf-c851-4571-a229-8ac4a5dd9455.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Size-Dependent Mating Success in Pink Bollworm moths: Insights into Mating Disruption Effects","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMale moths generally locate potential mates by detecting and following the pheromone plumes of females (Greenfield \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e1981\u003c/span\u003e; Card\u0026eacute; and Baker \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1984\u003c/span\u003e). This pheromone-mediated communication is crucial for reproduction in most moth species (Card\u0026eacute; \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Leveraging this process, the mating disruption technique, widely used in agricultural pest control, deploys synthetic pheromone components in fields to interfere with males' ability to locate females (Card\u0026eacute; \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Card\u0026eacute; and Minks \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Howse et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1998\u003c/span\u003e). The reduced ability of males to detect receptive females may be explained by several mechanisms, including competition with synthetic pheromone sources, masking of female pheromone plumes, sensory adaptation (reducing the responsiveness of olfactory receptors), and habituation (a central nervous system process that reduces behavioral responses to female pheromones) (Card\u0026eacute; and Minks \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Saunders 1997; Miller and Gut \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe efficacy of this technique depends on various factors, including the formulation and release method of the synthetic pheromone and environmental conditions such as canopy structure and wind direction shifts (Card\u0026eacute; \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Card\u0026eacute; and Minks \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Lykouressis et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). In some moth species, both sexes release pheromones attractive to the opposite sex (Landolt and Heath \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1990\u003c/span\u003e). In such species, disrupting only the male's orientation to the female may not fully prevent mating encounters (Card\u0026eacute; and Minks \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e). Additionally, the success of mating disruption can be reduced by migration of mated females into treated areas from external populations (Sexton and Il'ichev 2000; Saucke et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; R\u0026eacute;gni\u0026egrave;re et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). High pest densities can also diminish the effectiveness of this control method by increasing the likelihood of random encounters between males and females, amidst the synthetic pheromone plumes (Carpenter et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1982\u003c/span\u003e; Lance et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe success of mating disruption techniques varies among species. Notable successes include the oriental fruit moth (\u003cem\u003eGrapholita molesta\u003c/em\u003e), a key pest of stone and pome fruits (Rothschild \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e1975\u003c/span\u003e; Gentry et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1980\u003c/span\u003e; Vickers et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Il\u0026rsquo;chev et al. 2004; Kong et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2014\u003c/span\u003e); the tomato pinworm (\u003cem\u003eKeiferia lycopersicella\u003c/em\u003e), a major pest of tomatoes (Van Steenwyk et al. 1983; Wang et al. 1997; Schuster et al. 2000); the European grape moth (\u003cem\u003eEupoecilia ambiguella\u003c/em\u003e) and the grapevine moth (\u003cem\u003eLobesia botrana\u003c/em\u003e), both significant pests in European vineyards (Louis and Schirra \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Lioriatti et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Benelli et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Ricciardi et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2024\u003c/span\u003e); and the pink bollworm (\u003cem\u003ePectinophora gossypiella\u003c/em\u003e), a major pest of cotton worldwide (Gaston et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1977\u003c/span\u003e; Staten et al. 1987; Card\u0026eacute; \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Critchley et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Collar et al. 2002; Lykouressis et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2005\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe pink bollworm moth is a highly destructive cotton pest (Lukefahr and Martin \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e1963\u003c/span\u003e; Agarwal and Katiyar \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1979\u003c/span\u003e; Ingram \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e1980\u003c/span\u003e; Parmar and Pate 2016; Bhute et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Pink bollworm larvae primarily feed on developing cotton seeds, leading to premature boll opening or significant postharvest damage due to rotting (Parmar and Pate 2016; Sarwar \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The pink bollworm moth is challenging to manage with insecticides: Females predominantly lay eggs on the sutures or beneath the bracteoles at the base of bolls, and the larvae penetrate the bolls where they are protected from conventional insecticides (Lykouressis et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Sreenivas et al. 2021).\u003c/p\u003e\u003cp\u003eThe pink bollworm\u0026rsquo;s sex pheromone was identified in 1973 as a 1:1 blend of cis/cis (Z,Z) and cis/trans (Z,E) isomers of 7,11-hexadecadienyl acetate (Hummel et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1973\u003c/span\u003e), though deviations of this ratio have been documented (Haynes et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1984\u003c/span\u003e; Gonzalez-Karlsson et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Building on this discovery, studies in the late 1970s and early 1980s demonstrated promising results using 'Gossyplure', a synthetic formulation of this 1:1 blend, to disrupt the moths' mating (Gaston et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1977\u003c/span\u003e; Flint et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1978\u003c/span\u003e; Henneberry et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1981\u003c/span\u003e). Since then, mating disruption has been successfully used widely to control the pink bollworm moth in cotton fields (Brooks et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1979\u003c/span\u003e; Staten et al. 1987; Card\u0026eacute; and Minks \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Athanassiou et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Collar et al. 2002; Lykouressis et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). However, recent field evidence indicates that the mating disruption faces significant challenges in controlling the pink bollworm (Niv \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Harari and Sharon \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Research has suggested that mating disruption alone may not adequately control pink bollworm populations, highlighting the necessity to integrate other pest management strategies (Card\u0026eacute; and Minks \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Niv \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2000\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHere, we propose an evolutionary factor that may influence the efficacy of this important, environmentally friendly control method. The synthetic pheromone used for mating disruption of the pink bollworm moth consists of a 1:1 blend of its two components, reflecting the average pheromone composition of pink bollworm in field populations. However, larger and younger females in good condition tend to produce a higher ratio of Z,Z to Z,E isomers in their pheromone glands, which is recognized and preferred by males (Gonzalez-Karlsson et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). We hypothesized that the synthetic pheromone, which likely mirrors the emission ratios of the most abundant females in the population (i.e., medium-sized females), would predominantly affect the mating success of these females. A lesser impact on larger and smaller females is expected since their pheromone blend deviates from the 1: 1 ratio. Over evolutionary timescales, the higher mating success of large and small females under continuous mating disruption could lead to shifts in body size distribution within treated populations.\u003c/p\u003e\u003cp\u003eIf mating disruption disproportionately reduces the mating success of medium-sized females, this could lead to a shift in the mating proportions among other size classes in the population, with important implications for overall fecundity and the long-term efficacy of the technique. Since larger females in na\u0026iuml;ve populations exhibit higher fecundity than smaller females (Gonzalez-Karlsson et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), we hypothesized that this pattern would persist in field populations exposed to mating disruption. Therefore, an increase in the proportion of large females in treated fields could potentially undermine the overall efficacy of the mating disruption technique.\u003c/p\u003e\u003cp\u003eIn this study, we tested whether the mating success of females, across different size classes (large, medium, or small), differs under conditions of pheromone excess compared to na\u0026iuml;ve populations. We also examined the body size distribution of moths collected from mating disruption environments versus lab-reared moths, and investigated female fecundity based on their body size categories. By addressing these questions, this research provides the first evolutionary insights into how sexual selection affects the efficacy of mating disruption. Our findings also lay the foundation for developing improved strategies aiming to optimize this environmentally friendly pest control method for sustainable agricultural practices.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eBreeding and maintenance\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eTwo populations of pink bollworm moths were maintained in separate climate-controlled rooms at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, a 14:10 light-to-dark photoperiod and 60% relative humidity: (1) a na\u0026iuml;ve population collected before the implementation of mating disruption in cotton fields, never exposed to this method. These moths have been reared in the lab for approximately 40 years since their collection, and (2) a population established from larvae collected from cotton bolls in a field exposed to prolonged mating disruption. The latter group was reared in the laboratory for 10\u0026ndash;15 generations before being used in our experiments, maintained continuously under a pheromone-saturated environment, using synthetic pheromone ropes (1:1 ratio of the two components, Shin-Etsu, Japan). Larvae from both groups were reared on an artificial diet (Stonefly Heliothis Diet, Ward\u0026rsquo;s Science), and adults were provided with a 10% sugar solution ad libitum. Males and females were separated during the larval stage into two cages. Females were grouped by size during the pupae stage into three size categories: large (0.0170\u0026ndash;0.0220 g), medium (0.0120\u0026ndash;0.0160 g), and small (0.0070\u0026ndash;0.0110 g). Adult moths were separated daily into age-based (30 x 30 x 30 cm) screen cages.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eExperimental design\u003c/h3\u003e\n\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e\u003cb\u003eExperiment 1: The effect of mating disruption on female mating success in relation to their body size.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo test the hypothesis that mating disruption reduces the mating success of medium-sized females compared to na\u0026iuml;ve females, we conducted the following experiment: We placed 40 randomly sized males with 10 large, 10 medium, and 10 small females in a net cage. The insects were allowed to interact for 40 minutes during their mating window. Once a pair mated, they were removed, and the female\u0026rsquo;s size category (large, medium, or small) was identified using a pre-established visual scale. The scale was based on size of adult females that emerged from selected pupae of the different size groups (large, medium and small). This scale allowed us to reliably determine each female's category at the time of pair removal. The mated female was then replaced with another female of the same size category, while a new male was randomly selected to replace the removed male. This ensured a constant representation of all female size categories throughout the experiment. This experiment included the following three treatments:\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003col\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003eNa\u0026iuml;ve moths that have never been exposed to mating disruption, without synthetic pheromone during the experiment (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;17 trials).\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003eProlonged pheromone-exposed moths in the presence of synthetic pheromone during the experiment (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;15 trials).\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003eProlonged pheromone-exposed moths in the absence of synthetic pheromone during the experiment (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;15 trials).\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003c/ol\u003e\u003c/p\u003e\u003cp\u003eNote that na\u0026iuml;ve moths were not tested under mating disruption conditions. Previous studies showed that first exposure of na\u0026iuml;ve moths to synthetic pheromone causes a marked reduction in activity (Waner et al. 2025), making behavioral comparisons difficult and uninformative for the purpose of this study.\u003c/p\u003e\u003cp\u003eWe evaluated the proportion of medium-sized mated females across the three treatments by comparing the proportions of large versus medium-sized females and medium versus small-sized females.\u003c/p\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e\u003cem\u003eL\u003c/em\u003e\u0026minus;\u003cem\u003eM\u003c/em\u003e (Large minus Medium) represents the difference in proportions between large- and medium-sized mated females. Across all three treatments, the distribution of \u003cem\u003eL\u003c/em\u003e\u0026minus;\u003cem\u003eM\u003c/em\u003e was normal; therefore, t-tests were used to compare \u003cem\u003eL\u003c/em\u003e\u0026minus;\u003cem\u003eM\u003c/em\u003e between treatments. Similarly, \u003cem\u003eM\u003c/em\u003e\u0026minus;\u003cem\u003eS\u003c/em\u003e (Medium minus Small) represents the difference in proportions between medium- and small-sized mated females. Since the distribution of \u003cem\u003eM\u003c/em\u003e\u0026minus;\u003cem\u003eS\u003c/em\u003e was not normal for all treatments, comparisons of \u003cem\u003eM\u003c/em\u003e\u0026minus;\u003cem\u003eS\u003c/em\u003e were conducted using the non-parametric Mann-Whitney \u003cem\u003eU\u003c/em\u003e-test. Note that we were only interested in comparing medium-sized females to large- and small-sized females (but not large to small). Thus, we restricted our analysis to \u003cem\u003eL\u0026thinsp;\u0026minus;\u0026thinsp;M\u003c/em\u003e and \u003cem\u003eM\u0026thinsp;\u0026minus;\u0026thinsp;S\u003c/em\u003e to maximize statistical power.\u003c/p\u003e\u003cp\u003e\u003cb\u003eComparative Analysis 1: Comparing the size distribution of moths from na\u0026iuml;ve and pheromone-exposed populations.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo evaluate the differences in the size distribution between na\u0026iuml;ve moths and moths with prolonged exposure to mating disruption, we measured the weights of pupae randomly collected from the following three groups:\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003col\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eNa\u0026iuml;ve Population\u003c/b\u003e: Males (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;107) and females (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;97) from a population that has never been exposed to mating disruption.\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eField-Collected Population\u003c/b\u003e: Males (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;136) and females (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;143) collected as final-instar larvae from a cotton field treated with mating disruption.\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003e\u003cb\u003eLaboratory-Reared Field Population\u003c/b\u003e: Males (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;172) and females (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;183) from the same field-collected population as in group 2, reared for two generations under laboratory conditions similar to the na\u0026iuml;ve population but with the presence of synthetic pheromone (Shin-Etsu, Japan) in the rearing room. This approach controlled for the potential effect of laboratory diet on moth body size.\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003c/ol\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003estatistical analysis\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe weights of the 423 female and 415 male pupae were normally distributed. To assess potential weight differences based on origin, we conducted a one-way Welch's ANOVA for both female and male pupae, as this method is robust to unequal variances. In order to evaluate the overall significance of the weight differences between female and male pupae across all treatment groups, we employed Winer\u0026rsquo;s method of adding \u003cem\u003et\u003c/em\u003e-values (see Rosenthal, 1978).\u003c/p\u003e\u003cp\u003e\u003cb\u003eComparative Analysis 2: The impact of female size on fecundity in mating disruption-exposed populations.\u003c/b\u003e\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003ePrevious research has shown that larger na\u0026iuml;ve moths are more fecund than smaller na\u0026iuml;ve moths. To test whether female size affects fecundity in mating disruption-exposed females, we counted the number of eggs laid by groups of 10 females classified as large (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6 groups), medium (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6 groups), or small (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;5 groups). Each group was kept in a mating container with 15 randomly sized males. The excess of males aimed to ensure mating for all females. Egg-laying substrates were replaced twice weekly, and eggs were counted. Following the death of the females, dissections were performed to confirm the mating status of the females by the presence of at least one spermatophore.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eThe number of eggs did not have a normal distribution (Kolmogorov-Smirnov test: \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.037). Thus, a square-root transformation was applied for the one-way ANOVA analysis.\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cb\u003eExperiment 1: The effect of mating disruption on female mating success in relation to their body size.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eProportion of mated females:\u003c/p\u003e\u003cp\u003eIn the na\u0026iuml;ve population that was never exposed to synthetic pheromone, 47.41% of medium-sized females mated, compared to 39.26% of large females and 13.33% of small females. In contrast, in prolonged pheromone-exposed moths in the presence of synthetic pheromone, only 25.93% of medium-sized females mated, while 55.56% of large females and 18.52% of small females mated. For moths that were previously exposed to prolonged mating disruption but were not exposed to synthetic pheromone during the experiment, 43.09% of medium-sized females, 50.41% of large females, and 6.50% of small females successfully mated.\u003c/p\u003e\u003cp\u003eThere was no significant difference in \u003cem\u003eL\u003c/em\u003e\u0026minus;\u003cem\u003eM\u003c/em\u003e between the na\u0026iuml;ve population and the moths long exposed to mating disruption in the absence of synthetic pheromone (\u003cem\u003et\u003c/em\u003e\u003csub\u003e30\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.320, \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.751), with both groups showing similar mating percentages for large and medium-sized females. However, significant differences in \u003cem\u003eL\u003c/em\u003e\u0026minus;\u003cem\u003eM\u003c/em\u003e were found between the na\u0026iuml;ve population and the prolonged pheromone-exposed moths in the presence of the synthetic pheromone (\u003cem\u003et\u003c/em\u003e\u003csub\u003e30\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;2.838, \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.008), and between the prolonged pheromone-exposed moths in the presence vs. the absence of synthetic pheromone during the experiment (\u003cem\u003et\u003c/em\u003e\u003csub\u003e28\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;\u0026minus;2.703, \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.012), due to a lower proportion of mated medium-sized females in the presence of synthetic pheromone (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). There was no significant difference in \u003cem\u003eM\u003c/em\u003e\u0026minus;\u003cem\u003eS\u003c/em\u003e between the na\u0026iuml;ve population and the prolonged pheromone-exposed moths in the absence of synthetic pheromone (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.830). However, significant differences were observed between the na\u0026iuml;ve population vs. the prolonged pheromone-exposed moths with synthetic pheromone (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), as well as between the prolonged pheromone-exposed moths in the presence vs. the absence of synthetic pheromone during the experiment (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), again due to a lower proportion of medium-sized females mating in the presence of synthetic pheromone (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Percentage (average\u0026thinsp;\u0026plusmn;\u0026thinsp;se) of mated females in the three treatments: \u003cb\u003eA.\u003c/b\u003e Difference between large and medium sized (\u003cem\u003eL\u0026ndash;M\u003c/em\u003e) females. \u003cb\u003eB.\u003c/b\u003e Difference between medium and small sized \u003cem\u003e(M\u0026ndash;S\u003c/em\u003e) females.\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e\u003cb\u003eComparative Analysis 1: Comparing the size distribution of moths in na\u0026iuml;ve and pheromone-exposed populations.\u003c/b\u003e\u003c/p\u003e\u003cp\u003ePupae, males, and females originating from a cotton field under mating disruption and subsequently reared in the laboratory for two generations in the presence of synthetic pheromone were significantly heavier than na\u0026iuml;ve moths (both males and females) that had never been exposed to synthetic pheromone (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Across all groups, females were consistently larger than males (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001; Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eMultiple comparisons of weights by origin (Games-Howell method)\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMultiple comparisons\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFemales\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMales\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNa\u0026iuml;ve vs. Field-Collected\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNa\u0026iuml;ve vs. field, reared in the Laboratory\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eField-Collected vs. field, reared in the Laboratory\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.905\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.950\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e: Mean (\u0026plusmn;\u0026thinsp;SE) pupal weights of female and male moths from three population origins: na\u0026iuml;ve population, field-collected population (F0), and field-collected larvae reared in the laboratory for two generations under mating disruption conditions (F2). Orange bars represent females; blue bars represent males. Different Latin letters indicate statistically significant differences among female groups, while different Greek letters indicate significant differences among male groups.\u003c/p\u003e\n\u003ch3\u003eComparative Analysis 2:\u003c/h3\u003e\n\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e\u003cb\u003eThe impact of female size on fecundity in mating disruption-exposed populations.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eLarger females laid a significantly higher number of eggs than medium-size and small females. One-way ANOVA: \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001. Multiple comparisons: Small vs. Medium \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.087, Small vs. Large \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001, Medium vs. Large \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.039. (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eMating disruption is an environmentally friendly tactic used to control numerous moth pest species, including the pink bollworm moth in cotton fields (Card\u0026eacute; and Minks \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Staten et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). This species-specific approach minimizes harm to non-target organisms within the treated area. Instead of lethally targeting pests, it interferes with mating, thereby reducing reproductive success, lowering population levels, and ultimately decreasing crop damage (Mevada et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). While this technique has been largely effective against the pink bollworm, there remains room for improvement in ensuring its long-term efficacy.\u003c/p\u003e\u003cp\u003eAnalyses of the proportions of mated females across size categories revealed significant differences between pheromone-exposed moths, tested in the presence of synthetic pheromone, and na\u0026iuml;ve laboratory-reared moths. Similarly, significant size-related differences were also observed within the pheromone-exposed group when comparing trials conducted with and without synthetic pheromone. In contrast, no such differences were detected between laboratory-reared moths and pheromone-exposed moths when both were tested in the absence of synthetic pheromone.\u003c/p\u003e\u003cp\u003eThe proportion of small-mated females was consistently the lowest across all three experimental conditions. In the na\u0026iuml;ve population, which had never been exposed to mating disruption, medium-sized females mated at rates comparable to, or slightly higher than large females. Given that larger females generally have higher fecundity and are typically preferred by males (Gonzalez-Karlsson et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), this suggests that mating success is shaped not only by male preference for high-quality mates but also by female selectivity. Larger, higher-quality pink bollworm females tend to be more selective, while smaller females are less preferred by males (Waner Rips, submitted). As a result, medium-sized females, though not the highest-quality mates, may achieve relatively high mating success by being less selective than large females and more attractive than small ones. Interestingly, the proportion of mated medium-sized females dropped sharply under pheromone-treated conditions, influencing the mating proportions of both large and small females. These shifts are reflected in the differences in the mating rate between large and medium females (L\u0026thinsp;\u0026minus;\u0026thinsp;M), and between medium and small females (M\u0026thinsp;\u0026minus;\u0026thinsp;S) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). These findings suggest that the standardized 1:1 ratio of pheromone components used in mating disruption exerts size-dependent effects, disproportionately reducing the mating success of medium-sized females.\u003c/p\u003e\u003cp\u003eA previous study (Gonzalez-Karlsson et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) showed that large females produce a higher proportion of the Z,Z isomer, while small females produce more of the Z,E isomer in their sex pheromone blends. Both deviate from the 1:1 (Z,Z):(Z,E) ratio used in the commercial mating disruption formulation (Shin-Etsu, Japan). Although the pheromone blends of females in our study were not directly analyzed, large and small females likely retained their respective isomer biases. Medium-sized females, being the most prevalent in the population, likely emitted blends closer to the synthetic 1:1 ratio, rendering them more susceptible to pheromone masking. This may explain their reduced mating success under disrupted conditions, while the less common blends of large and small females were less affected.\u003c/p\u003e\u003cp\u003eAnother notable finding of this study is that moths collected from pheromone-treated cotton fields were significantly larger than laboratory-reared moths. This size difference persisted even after two generations of rearing under identical laboratory conditions.\u003c/p\u003e\u003cp\u003eBoth sexual selection and natural selection (including environmental changes) generally influence the body size of insects; Larger female insects typically achieve greater reproductive success (Wickman and Karlsson \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e1989\u003c/span\u003e), thus being favored by sexual selection processes. This can drive a shift toward larger female body size, particularly in species where larger females are strongly preferred by males (Andersson and Iwasa \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1996\u003c/span\u003e). Similarly, sexual selection tends to favor larger males, as they often gain a competitive advantage in securing mates, thereby achieving more copulations (Alcock \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Serrano-Meneses et al. \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). The reproductive advantage of large males and females is well-documented across taxa (Olsson et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Byrne and Rice \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2006\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAt the same time, natural selection often counterbalances this trend by favoring intermediate phenotypes, such as average body size, over extreme ones (Blanckenhorn \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). Selection for intermediate body size, a form of stabilizing selection, occurs when extreme sizes come with fitness disadvantages. Smaller females often have reduced fecundity and energy reserves, while larger females may experience increased energy demands, higher developmental costs, greater predation risk (Kullberg et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Neems et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Blanckenhorn \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Covas et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2002\u003c/span\u003e), or an overall higher mortality rate (Xu and Wang \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Environmental changes can shift body size distributions in insects, as demonstrated for climate-driven trends in various species (Brose et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn our study, we propose that the synthetic pheromone may weaken stabilizing selection in disrupted populations by disproportionately reducing the mating success of medium-sized females. Since males rely on pheromonal cues to locate mates, and medium-sized females are more likely to emit blends resembling the synthetic 1:1 ratio used in disruption, their signals may be more effectively masked. As a result, large females, whose pheromone blends deviate from the synthetic standard, may achieve higher mating rates, potentially shifting body size distributions in the population. Furthermore, the masking of medium-sized females may increase mating opportunities for smaller, less-preferred females, as males lower their selectivity when mate location becomes more challenging. Consistent with this, we observed a slight increase in the mating success of small females under pheromone-saturated conditions. However, because small females are generally less preferred, this shift likely contributes less to selection pressures than the increased mating of large females.\u003c/p\u003e\u003cp\u003eEnvironmental factors such as diet or population density may also affect body size. Our na\u0026iuml;ve population had been reared in the laboratory for ~\u0026thinsp;40 years, which could have influenced female size, although previous studies suggest long-term lab rearing has little effect on lepidopteran body size (Hoffmann and Ross \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Still, we cannot exclude the possibility that it also contributed to the smaller sizes observed in na\u0026iuml;ve moths compared with field-collected moths. To specifically address the potential influence of laboratory diet, one of the main factors that could affect lab moths\u0026rsquo; size, we reared a subset of the field-collected population in the laboratory for two additional generations under controlled conditions, with synthetic pheromones present. The pupae from this second generation were also significantly larger than those from the na\u0026iuml;ve population, and their size did not differ from that of pupae collected directly from treated fields.\u003c/p\u003e\u003cp\u003eInsect body size is influenced by genetic and environmental factors and plays a critical role in determining fitness (Nijhout \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Edgar \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Beukeboom \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Nijhout et al. 2003). Although specific data disentangling the genetic and environmental influences on body size in the pink bollworm is unavailable, both factors are expected to play a role. If this pattern persists across generations, it could have contributed to an increase in the average size of moths in treated areas. Female size in insects is often a reliable indicator of fecundity (Honěk \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1993\u003c/span\u003e). Large female pink bollworm moths that have never been exposed to mating disruption are more fecund than smaller moths (Gonzalez-Karlsson et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). We re-tested this relationship in moths from a field treated with mating disruption for many generations. Similarly, we found that larger females were significantly more fecund than medium-sized and small females, as measured by the total number of eggs laid over their lifetimes.\u003c/p\u003e\u003cp\u003eA higher proportion of larger females, coupled with their higher fecundity, in areas subjected to mating disruption, raises questions about the long-term efficacy of this method. If larger females consistently contribute more offspring, this could influence population structure dynamics and potentially reduce the effectiveness of mating disruption over time. To address this challenge, exploring and applying alternative ratios of sex pheromone isomers could enhance the efficacy of this promising and environmentally friendly approach. Further research into the interplay between female size, fecundity, and pheromone communication under mating disruption conditions may provide valuable insights, contributing to the refinement of this method for sustainable pest management.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eEthical Note\u003c/h2\u003e\u003cp\u003eThe pink bollworm moth (\u003cem\u003ePectinophora gossypiella\u003c/em\u003e) is a well-known agricultural pest. Consequently, research involving this species typically does not require specific ethical permits. Nonetheless, all experimental procedures were conducted in adherence to ethical standards. The insects were reared in climate-controlled rooms maintained at a constant temperature of 25\u0026deg;C and were consistently provided with appropriate food corresponding to their developmental stages. Throughout the study, the moths were handled gently and with care. After mating trials, they were released. At no point were the insects harmed or subjected to detrimental conditions.\u003c/p\u003e\u003c/div\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eSWR conceived the research questions and designed the experiments with the help of AH and MM. SWR conducted the experiments, analyzed the data, and wrote the manuscript. AH and MM assisted in experiment design, conducted the experiments, and reviewed the manuscript. UM contributed to the statistical analysis and reviewed the manuscript. OK took part in conceiving the research questions and revising the manuscript. All authors read and approved the final version of the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eWe thank Arnon Lotem and Victoria Soroker for their valuable advice and insightful discussions that greatly improved this study. We are also deeply grateful to Prof. Ariel Chipman for his continuous support and thoughtful advice whenever needed. Our thanks extend to Eyal Halon, Michael Davidivitz, Nikolay Meltser, and Aya Rafael Cohen for their dedicated technical assistance and field support. This research was partially supported by the Israel Science Foundation (Grant No. 1826/20), the British Friends of the Hebrew University, and the Yael Pitoun Scholarship.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAgarwal RA, Katiyar KN (1979) An estimate of losses of seed kapas and seed due to bollworms on cotton in India. Indian J Entomol 41:143\u0026ndash;148\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlcock J (1994) Body size and its effect on male-male competition in \u003cem\u003eHylaeus alcyoneus\u003c/em\u003e (Hymenoptera: Colletidae). J Insect Behav 8:149\u0026ndash;159 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF01988901\u003c/span\u003e\u003cspan address=\"10.1007/BF01988901\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAndersson M, Iwasa Y (1996) Sexual selection. Trends Ecol Evol 11:53\u0026ndash;58 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0169-5347(96)81042-1\u003c/span\u003e\u003cspan address=\"10.1016/0169-5347(96)81042-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAthanassiou CG, Buchelos CT, Kavallieratos NG, Barbetaki AE (2002) Evaluation of the mating disruption method for the control of the pink bollworm \u003cem\u003ePectinophora gossypiella\u003c/em\u003e (Saund.) (Lepidoptera: Gelechiidae) and comparison of this method with insecticidal treatments. IOBC WPRS Bull 25\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBenelli G, Lucchi A, Anfora G, Bagnoli B, Botton M, Campos-Herrera R et al. (2023) European grapevine moth, \u003cem\u003eLobesia botrana\u003c/em\u003e. Part II: Prevention and management. Entomologia Generalis 43(2):281\u0026ndash;304 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1127/entomologia/2023/1947\u003c/span\u003e\u003cspan address=\"10.1127/entomologia/2023/1947\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBeukeboom LW (2018) Size matters in insects: An introduction. Entomologia Experimentalis et Applicata 166(1):2\u0026ndash;3 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/eea.12646\u003c/span\u003e\u003cspan address=\"10.1111/eea.12646\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBhute NK, Patil CS, Deshmukh KV, Wagh RS, Medhe NK (2023) Pink bollworm \u003cem\u003ePectinophora gossypiella\u003c/em\u003e (Saunders), a destructive pest of cotton: a review. Pharma Innovation J 12(3):2036\u0026ndash;2042\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBlanckenhorn WU (2000) The evolution of body size: what keeps organisms small? Quart Rev Biol 75(4):385\u0026ndash;407 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1086/393620\u003c/span\u003e\u003cspan address=\"10.1086/393620\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBrooks TW, Doane CC, Staten RT (1979) Experience with the first commercial pheromone communication disruptor for suppression of an agricultural insect pest. \u003cem\u003eIn: Ritter FJ (ed) Chemical Ecology: Odour Communication in Animals\u003c/em\u003e. Elsevier/North-Holland Biomedical Press, Amsterdam, pp 375\u0026ndash;388\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBrose U, Dunne JA, Montoya JM, Petchey OL, Schneider FD, Jacob U (2012) Climate change in size-structured ecosystems. Phil Trans R Soc B 367:2903\u0026ndash;2912 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1098/rstb.2012.0232\u003c/span\u003e\u003cspan address=\"10.1098/rstb.2012.0232\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eByrne PG, Rice WR (2006) Evidence for adaptive male mate choice in the fruit fly \u003cem\u003eDrosophila melanogaster\u003c/em\u003e. Proc R Soc B 273:917\u0026ndash;922 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1098/rspb.2005.3372\u003c/span\u003e\u003cspan address=\"10.1098/rspb.2005.3372\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCard\u0026eacute; RT, Baker TC (1984) Sexual communication with pheromones. \u003cem\u003eIn: Bell WJ, Card\u0026eacute; RT (eds) Chemical Ecology of Insects\u003c/em\u003e. Springer US, Boston MA, pp 355\u0026ndash;383\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCard\u0026eacute; RT (1990) Principles of mating disruption. \u003cem\u003eIn: Ridgway RL, Silverstein RM, Inscoe MN (eds) Behavior-Modifying Chemicals for Pest Management: Applications of Pheromones and Other Attractants\u003c/em\u003e. Marcel Dekker, New York, pp 47\u0026ndash;71\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCard\u0026eacute; RT, Minks AK (1995) Control of moth pests by mating disruption: successes and constraints. Annu Rev Entomol 40:559\u0026ndash;585 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1146/annurev.en.40.010195.003015\u003c/span\u003e\u003cspan address=\"10.1146/annurev.en.40.010195.003015\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCard\u0026eacute; RT (2016) Moth navigation along pheromone plumes. \u003cem\u003eIn: Allison JD \u0026amp; Card\u0026eacute; RT (eds) Pheromone Communication in Moths: Evolution, Behavior, and Application\u003c/em\u003e. University of California Press, Berkeley, pp 173\u0026ndash;190 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1525/9780520964433-012\u003c/span\u003e\u003cspan address=\"10.1525/9780520964433-012\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCarpenter JE, Sparks AN, Gueldner RC (1982) Effects of moth population density and pheromone concentration on mating disruption of the corn earworm in large screened cages. J Econ Entomol 75:333\u0026ndash;336 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/jee/75.2.333\u003c/span\u003e\u003cspan address=\"10.1093/jee/75.2.333\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCollar Urquijo JL, G\u0026aacute;lvez Manzano V (2002) Mating disruption strategies for the control of pink bollworm (\u003cem\u003ePectinophora gossypiella\u003c/em\u003e Saunders) in cotton fields. Bolet\u0026iacute;n Sanid Veg Plagas 28:193\u0026ndash;198\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCovas R, Brown CR, Anderson MD, Brown MB (2002) Stabilizing selection on body mass in the sociable weaver \u003cem\u003ePhiletairus socius\u003c/em\u003e. Proc R Soc B 269:1905\u0026ndash;1909 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1098/rspb.2002.2106\u003c/span\u003e\u003cspan address=\"10.1098/rspb.2002.2106\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCritchley BR, Chamberlain DJ, Campion DG, Attique MR, Ali M, Ghaffar A (1991) Integrated use of pink bollworm pheromone formulations and selected conventional insecticides for the control of the cotton pest complex in Pakistan. Bull Entomol Res 81:371\u0026ndash;378\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eEdgar BA (2006) How flies get their size: genetics meets physiology. Nat Rev Genet 7:907\u0026ndash;916 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/nrg1989\u003c/span\u003e\u003cspan address=\"10.1038/nrg1989\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFlint HM, Butler LI, McDonough LM, Smith RL, Forey DE (1978) Pink bollworm: response to various emission rates of gossyplure in the field. Environ Entomol 7:57\u0026ndash;61 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/ee/7.1.57\u003c/span\u003e\u003cspan address=\"10.1093/ee/7.1.57\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGaston LK, Kaae RS, Shorey HH, Sellers D (1977) Controlling the pink bollworm by disrupting sex pheromone communication between adult moths. Science 196:904\u0026ndash;905 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1126/science.870967\u003c/span\u003e\u003cspan address=\"10.1126/science.870967\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGentry CR, Bierl-Leonhardt BA, Blythe JL, Plimmer JR (1980) Air permeation tests with orfralure for reduction in trap catch of oriental fruit moths. J Chem Ecol 6:185\u0026ndash;192\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGonzalez-Karlsson A, Golov Y, Steinitz H, Moncaz A, Halon E, Horowitz AR, Goldenberg I, Gurka R, Liberzon A, Soroker V, Jurenka R (2021) Males perceive honest information from female-released sex pheromone in a moth. Behav Ecol 32:1127\u0026ndash;1137 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/beheco/arab073\u003c/span\u003e\u003cspan address=\"10.1093/beheco/arab073\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGreenfield MD (1981) Moth sex pheromones: an evolutionary perspective. Fla Entomol 64:4\u0026ndash;17 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2307/3494597\u003c/span\u003e\u003cspan address=\"10.2307/3494597\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHarari A, Sharon R (2022) The contemporary and prospective risks of resistance to the mating disruption method in moths. Entomol Gen 42(2):275\u0026ndash;288 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1127/entomologia/2021/1275\u003c/span\u003e\u003cspan address=\"10.1127/entomologia/2021/1275\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHaynes KF, Gaston LK, Pope MM, Baker TC (1984) Potential for evolution of resistance to pheromones: interindividual and interpopulational variation in the chemical communication system of the pink bollworm moth. J Chem Ecol 10:1551\u0026ndash;1565 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF00988424\u003c/span\u003e\u003cspan address=\"10.1007/BF00988424\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHoněk A (1993) Intraspecific variation in body size and fecundity in insects: a general relationship. Oikos 66(3):483\u0026ndash;492 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2307/3544943\u003c/span\u003e\u003cspan address=\"10.2307/3544943\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHenneberry TJ, Gillespie JM, Bariola LA, Flint HM, Lingren PD, Kydonieus AF (1981) Gossyplure in laminated plastic formulations for mating disruption and pink bollworm control. J Econ Entomol 74(4):376\u0026ndash;381 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/jee/74.4.376\u003c/span\u003e\u003cspan address=\"10.1093/jee/74.4.376\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHoffmann AA, Ross PA (2018) Rates and patterns of laboratory adaptation in (mostly) insects. J Econ Entomol 111(2):501\u0026ndash;509 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/jee/toy024\u003c/span\u003e\u003cspan address=\"10.1093/jee/toy024\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHowse PE, Stevens IDR, Jones OT (1998) Mating disruption. In: Howse PE, Stevens IDR, Jones OT (eds) Insect Pheromones and Their Use in Pest Management. Springer, Dordrecht, pp 314\u0026ndash;344 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-94-011-5344-7_12\u003c/span\u003e\u003cspan address=\"10.1007/978-94-011-5344-7_12\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHummel HE, Gaston LK, Shorey HH, Kaae RS, Byrne KJ, Silverstein RM (1973) Clarification of the chemical status of the pink bollworm sex pheromone. Science 181(4102):873\u0026ndash;875 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1126/science.181.4102.873\u003c/span\u003e\u003cspan address=\"10.1126/science.181.4102.873\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eIngram WR (1980) Studies of the pink bollworm, Pectinophora gossypiella, on sea island cotton in Barbados. Int J Pest Manag 26(2):118\u0026ndash;137 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.tandfonline.com/doi/abs/10.1080/09670878009414384\u003c/span\u003e\u003cspan address=\"https://www.tandfonline.com/doi/abs/10.1080/09670878009414384\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKong WN, Li J, Fan RJ, Li SC, Ma RY (2014) Sex-pheromone-mediated mating disruption technology for the oriental fruit moth, \u003cem\u003eGrapholita molesta\u003c/em\u003e (Busck) (Lepidoptera: Tortricidae): overview and prospects. Psyche A J Entomol 2014:253924 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1155/2014/253924\u003c/span\u003e\u003cspan address=\"10.1155/2014/253924\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKullberg C, Fransson T, Jakobsson S (1996) Impaired predator evasion in fat blackcaps (\u003cem\u003eSylvia atricapilla\u003c/em\u003e). Proc R Soc B 263(1377):1671\u0026ndash;1675 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1098/rspb.1996.0244\u003c/span\u003e\u003cspan address=\"10.1098/rspb.1996.0244\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLandolt PJ, Heath RR (1990) Sexual role reversal in mate-finding strategies of the cabbage looper moth. Science 249(4972):1026\u0026ndash;1028 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1126/science.249.4972.1026\u003c/span\u003e\u003cspan address=\"10.1126/science.249.4972.1026\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLance DR, Leonard DS, Mastro VC, Walters ML (2016) Mating disruption as a suppression tactic in programs targeting regulated lepidopteran pests in US. J Chem Ecol 42(7):590\u0026ndash;605 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10886-016-0732-9\u003c/span\u003e\u003cspan address=\"10.1007/s10886-016-0732-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLouis F, Schirra KJ (2001) Mating disruption of \u003cem\u003eLobesia botrana\u003c/em\u003e (Lepidoptera: Tortricidae) in vineyards with very high population densities. IOBC WPRS Bull 24(2):75\u0026ndash;80\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eIoriatti CL, Bagnoli BR, Lucchi AN, Veronelli VI (2004) Vine moths control by mating disruption in Italy: results and future prospects. Redia 87:117\u0026ndash;128\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eIl\u0026rsquo;ichev AL, Williams DG, Milner AD (2004) Mating disruption barriers in pome fruit for improved control of oriental fruit moth \u003cem\u003eGrapholita molesta\u003c/em\u003e (Busck) in stone fruit under mating disruption. J Appl Entomol 128(2):126\u0026ndash;132 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1439-0418.2004.00822.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1439-0418.2004.00822.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLukefahr MJ, Martin DF (1963) Evaluation of damage to lint and seed of cotton caused by the pink bollworm. J Econ Entomol 56(5):710\u0026ndash;713\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLykouressis D, Perdikis D, Samartzis D, Fantinou A, Toutouzas S (2005) Management of the pink bollworm \u003cem\u003ePectinophora gossypiella\u003c/em\u003e (Saunders) by mating disruption in cotton fields. Crop Prot 24(2):177\u0026ndash;183 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cropro.2004.07.007\u003c/span\u003e\u003cspan address=\"10.1016/j.cropro.2004.07.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMevada RR, Sisodiya DB, Parmar RG, Prajapati DR (2023) Mating disruption: an ecological step towards sustainable pest management. J Eco-Friendly Agric 18(1):144\u0026ndash;150 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5958/2582-2683.2023.00025.4\u003c/span\u003e\u003cspan address=\"10.5958/2582-2683.2023.00025.4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMiller JR, Gut LJ (2015) Mating disruption for the 21st century: matching technology with mechanism. Environ Entomol 44(3):427\u0026ndash;453 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/ee/nvv052\u003c/span\u003e\u003cspan address=\"10.1093/ee/nvv052\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNeems RM, Lazarus J, McAdllan AJ (1998) Lifetime reproductive success in a swarming midge: trade-offs and stabilizing selection for male body size. Behav Ecol 9(3):279\u0026ndash;286 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/beheco/9.3.279\u003c/span\u003e\u003cspan address=\"10.1093/beheco/9.3.279\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNijhout HF (2003) The control of body size in insects. Dev Biol 261(1):1\u0026ndash;9 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0012-1606(03)00276-8\u003c/span\u003e\u003cspan address=\"10.1016/S0012-1606(03)00276-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNijhout HF, Riddiford LM, Mirth CK, Shingleton A, Suzuki Y, Callier V (2014) The developmental control of size in insects. WIREs Dev Biol 3(1):113\u0026ndash;134 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/wdev.124\u003c/span\u003e\u003cspan address=\"10.1002/wdev.124\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNiv A (2000) Use of pheromones for pink bollworm (\u003cem\u003ePectinophora gossypiella\u003c/em\u003e, Saunders) mating disruption in Israel. In: Proceedings of the World Cotton Research Conference, Vol 2, pp 6\u0026ndash;12\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eOlsson M, Shine R, Wapstra E, Ujvari B, Madsen T (2002) Sexual dimorphism in lizard body shape: the roles of sexual selection and fecundity selection. Evolution 56(7):1538\u0026ndash;1542 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.0014-3820.2002.tb01464.x\u003c/span\u003e\u003cspan address=\"10.1111/j.0014-3820.2002.tb01464.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eParmar VR, Patel CC (2016) Pink bollworm: a notorious pest of cotton: a review. AGRES Int e-J 5(2):88\u0026ndash;97\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eR\u0026eacute;gni\u0026egrave;re J, Delisle J, Dupont A, Trudel R (2019) The impact of moth migration on apparent fecundity overwhelms mating disruption as a method to manage spruce budworm populations. Forests 10(9):775 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/f10090775\u003c/span\u003e\u003cspan address=\"10.3390/f10090775\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRicciardi R, Benelli G, Di Giovanni F, Lucchi A (2024) The European grape berry moth, \u003cem\u003eEupoecilia ambiguella\u003c/em\u003e (Lepidoptera: Tortricidae): Current knowledge and management challenges. Crop Prot p 106641 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cropro.2024.106641\u003c/span\u003e\u003cspan address=\"10.1016/j.cropro.2024.106641\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRothschild GHL (1975) Control of oriental fruit moth (\u003cem\u003eCydia molesta\u003c/em\u003e (Busck)) (Lepidoptera: Tortricidae) with synthetic female pheromone. Bull Entomol Res 65(3):473\u0026ndash;490 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/S0007485300006143\u003c/span\u003e\u003cspan address=\"10.1017/S0007485300006143\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSaucke H, Balasus A, Finckh MR, Formowitz B, Schmidt R, Kratt A (2014) Mating disruption of pea moth (\u003cem\u003eCydia nigricana\u003c/em\u003e) in organic peas (\u003cem\u003ePisum sativum\u003c/em\u003e). Entomol Exp Appl 150(3):199\u0026ndash;207 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/eea.12153\u003c/span\u003e\u003cspan address=\"10.1111/eea.12153\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSarwar M (2017) Biological parameters of pink bollworm \u003cem\u003ePectinophora gossypiella\u003c/em\u003e (Saunders) (Lepidoptera: Gelechiidae): a looming threat for cotton and its eradication opportunity. Int J Res Agric For 4(7):25\u0026ndash;36\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSerrano-Meneses MA, C\u0026oacute;rdoba-Aguilar A, M\u0026eacute;ndez V, Layen SJ, Sz\u0026eacute;kely T (2007) Sexual size dimorphism in the American rubyspot: male body size predicts male competition and mating success. Anim Behav 73(6):987\u0026ndash;997 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.anbehav.2006.08.01\u003c/span\u003e\u003cspan address=\"10.1016/j.anbehav.2006.08.01\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStaten RT, El-Lissy O, Antilla L (1997) Successful area-wide program to control pink bollworm by mating disruption. In: Card\u0026eacute; RT, Minks AK (eds) \u003cem\u003eInsect Pheromone Research: New Directions\u003c/em\u003e. Springer US, Boston MA, pp 383\u0026ndash;396\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVickers RA, Rothschild GHL, Jones EL (1985) Control of the oriental fruit moth, \u003cem\u003eCydia molesta\u003c/em\u003e (Busck) (Lepidoptera: Tortricidae), at a district level by mating disruption with synthetic female pheromone. Bull Entomol Res 75(4):625\u0026ndash;634 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/S0007485300015881\u003c/span\u003e\u003cspan address=\"10.1017/S0007485300015881\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWickman PO, Karlsson B (1989) Abdomen size, body size and the reproductive effort of insects. Oikos 56(2):209\u0026ndash;214 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2307/3565338\u003c/span\u003e\u003cspan address=\"10.2307/3565338\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eXu J, Wang Q (2013) Trade-off between adult body size and juvenile survival: an experimental test of parental effects in the Mediterranean flour moth. Aust J Entomol 52:403\u0026ndash;406 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/aen.12038\u003c/span\u003e\u003cspan address=\"10.1111/aen.12038\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-7842351/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7842351/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMating disruption is an environmentally friendly pest management technique that interferes with pheromone communication in moths, disrupting males\u0026rsquo; ability to locate females, thereby reducing pest populations in agricultural systems. This study explores how mating disruption affects mating success and size distribution in the pink bollworm (\u003cem\u003ePectinophora gossypiella\u003c/em\u003e), a major cotton pest worldwide. The species pheromone normally consists of a 1:1 ratio of Z,Z- and Z, E-7,11-hexadecadienyl acetate isomers, although larger females emit a slightly higher proportion of the Z,Z isomer. The synthetic pheromone used for disruption matches the average blend ratio (1:1 ZZ:ZE), assumed to represent the most abundant medium-sized females. We hypothesized that in pheromone-saturated environments, males would face difficulties locating medium-sized females whose pheromone resembles the synthetic blend. Consequently, we predicted reduced mating success for medium-sized females, potentially shifting body size distributions in the population. To test this, we compared the mating success of large, medium, and small females from a laboratory \u0026ldquo;na\u0026iuml;ve\u0026rdquo; population, never exposed to synthetic pheromone, and from a field population exposed to pheromone for many generations, with and without synthetic pheromone. We also measured pupal weights to assess body size differences. Results showed that medium-sized females experienced significantly reduced mating success in the presence of synthetic pheromone. Additionally, moths from mating disruption\u0026ndash;treated fields were, on average, larger than those from the lab. Because larger females are more fecund, laying substantially more eggs than medium- or small-sized females, their increased mating success could partially counteract the suppression effect of the mating disruption strategy.\u003c/p\u003e","manuscriptTitle":"Size-Dependent Mating Success in Pink Bollworm moths: Insights into Mating Disruption Effects","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-27 19:29:29","doi":"10.21203/rs.3.rs-7842351/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"dcba40bc-cd64-4b43-b55c-2e7cb0612a07","owner":[],"postedDate":"November 27th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-02T09:57:17+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-27 19:29:29","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7842351","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7842351","identity":"rs-7842351","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}