Fitness advantage from nuptial gifts in female fireflies

Abstract 1. In many insects, males provide nuptial gifts to females in the form of spermatophores, sperm-containing structures produced by male accessory glands.
2. The work reported here examined the influence of both spermatophore number and spermatophore size on female reproductive output in two related firefly beetles, Photinus ignitus and Ellychnia corrusca (Coleoptera: Lampyridae). Based on differences in adult diet, male spermatophores were predicted to increase female reproductive output to a greater extent in P. ignitus than in E. corrusca .
3. Female fecundity was significantly higher in triply mated females than in singly mated females in both species, with no difference between mating treatments in female lifespan or egg hatching success. No effects of second male spermatophore size on fecundity, lifespan, or egg hatching success were detected in either species.
4. These results suggest a direct fitness advantage from multiple mating for females in both species, although enhanced fecundity may be due either to allocation of spermatophore nutrients to eggs or to other substances transferred within the spermatophore acting as oviposition stimulants.     

Multiple mating of male and female codling moth (Lepidoptera: Tortricidae) in apple orchards treated with sex pheromone

Studies were conducted with codling moth, Cydia pomonella L., to evaluate the mating status of male and female moths in apple, Malus domestica (Borkhausen), orchards treated with and without sex pheromone dispensers. Laboratory studies first examined the effect of multiple mating of male and female moths on female fecundity and egg fertility. Females that had mated three times had a significantly higher fecundity than singly mated moths. Sequential mating by male moths had no effect on the fecundity of female moths or egg fertility. However, male moth age did impact female fecundity, with significantly fewer eggs laid after mating with virgin 1- versus 3-d-old males. The mean size of the first spermatophore transferred by males was significantly larger than all subsequent spermatophores. Classifying spermatophores based on size was used in field sampling to categorize the mating status of the female's partner. The proportion of mated females with small spermatophores (partner had previously mated) was significantly higher in treated versus untreated orchards. The proportion of female moths caught in traps baited with pear ester that were virgin was low (相似文献   

Effects of male mating interval on spermatophore formation,transfer, and subsequent female receptivity and fecundity in the sorghum plant bug,Stenotus rubrovittatus

Males of the sorghum plant bug, Stenotus rubrovittatus (Matsumura) (Heteroptera: Miridae), transfer a spermatophore to females during copulation. After a 1‐day interval between the first and second copulation, males transferred both sperm and a spermatophore to females during the second copulation. However, when male mating interval was <1 h, they transferred sperm but no spermatophores to females during the second copulation. Therefore, the male mating interval probably produces two types of mated females, those with and those without a spermatophore. Mated females of S. rubrovittatus do not remate for at least 3 days after mating, even when courted, and lay more eggs than virgin females at the beginning of the oviposition period. The effects of spermatophores on female sexual receptivity and fecundity were examined using mated females with or without a spermatophore. Only one of the 40 (2.5%) mated females with a spermatophore remated, whereas 10 of the 26 (38.5%) without a spermatophore remated. Furthermore, mated females with a spermatophore laid more eggs than those without a spermatophore. These results suggest that spermatophores participate in reducing female sexual receptivity and enhancing female fecundity in S. rubrovittatus.     

Male larval nutrition in Choristoneura rosaceana (Lepidoptera: Tortricidae): an important factor in reproductive success

This study examines how Choristoneura rosaceana male quality, as determined by larval diet, age and mating history, affects the reproductive success of both sexes. While the size of the spermatophore produced at first mating increased linearly with male age, the frequency of mating was significantly higher for middle-aged males (2–4 days old) than younger (0–2 days old) or older (6–8 days old) individuals, when both sexes were fed on artificial diet. However, the duration of copulation was longer in couples with older than younger males. The observed age-related changes in spermatophore size had no significant effect on female longevity, fecundity or fertility, suggesting no direct relationship between male investment and spermatophore size under these experimental conditions. Different larval food sources (artificial diet, maple and hazelnut) did not affect the proportion of 2-day-old virgin males that mated; however, the proportion that remated was significantly higher for males reared on high-quality food (maple and artificial diet) than those on hazelnut, a poorer food source. There was a 5-fold decline in spermatophore size between the first and second matings on all diets, but female reproductive output was reduced by only 25%. In contrast, while the first spermatophore produced by males on hazelnut was 1.5 times smaller than those produced on maple and artificial diet, the fecundity of their mates was 40% less than those mated with high-quality virgin males. These results provide additional support to the idea that spermatophore size is not a valuable indicator of male quality. Most tethered females placed in the field during the first flight period mated with virgin males (based on the size of the spermatophore), suggesting that female choice exists in this species. These results are discussed in relation to the incidence of polyandry in naturally occurring populations of Choristoneura and the potential use of size and/or chemical cues by females to assess male quality.     

Spermatophore and Sperm Allocation in Males of the Monandrous Butterfly Pararge aegeria: the Female’s Perspective

In insects, spermatophore production represents a non‐trivial cost to a male. Non‐virgin males have been shown to produce small spermatophores at subsequent matings. Particularly in monandrous species, it may be an issue to receive a sufficiently large spermatophore at the first and typically only mating. Females of the monandrous Speckled wood butterfly Pararge aegeria (L.) produce fewer offspring after mating with a non‐virgin male. After mating, females spend all their active time selecting oviposition sites and typically ignore other males. Here, we show that females did not discriminate between a virgin male and a recently mated male in our laboratory experiments. We demonstrate that the number of eupyrene sperm bundles relative to spermatophore mass differed with subsequent male matings. Males transferred a significantly smaller spermatophore after the first copulation, but the spermatophore mass did not decrease further with subsequent matings. However, the number of eupyrene sperm bundles decreased linearly. Therefore, there was proportionally more eupyrene sperm in the male’s second spermatophore compared with the first and the later spermatophores. Such a pattern has been shown in polyandrous species. Hence, it suggests that differences in sperm allocation strategy between polyandrous and monandrous butterflies may be quantitative rather than qualitative. There was also a tendency for females that had mated with a recently mated male to have higher propensity to remate than did females that had mated with a virgin male. We discuss the results relative to the mating system in P. aegeria, including female remating opportunities in the field and male mate‐locating behaviour.     

Quantifying rates of random mating in western corn rootworm emerging from Cry3Bb1‐expressing and refuge maize in field cages

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is the most significant pest of field maize, Zea mays L. (Poaceae), in the USA. Maize plants expressing Bt toxins targeting the corn rootworm complex have been widely adopted and are the primary insecticidal control measure for this pest in North America. Insect resistance management tactics using various refuge structures have been adopted to ensure Bt products will retain durability. An assumption of the refuge strategy is that males and females emerging from Bt and refuge plantings mate randomly; this has not been tested in the field. We conducted cage studies using field populations of WCR in Indiana, USA, to generate empirical field data on mating rates between beetles emerging from Cry3Bb1‐expressing Bt and refuge maize plants. Two refuge configurations were tested; all refuge plants were labeled using the stable isotope ¹⁵N. This mark persists in adult beetles after eclosion, allowing for collection and analysis of isotopic ratios of all beetles. Additional data collected included adult emergence rates, timing and sex ratios for each of the treatments, and head capsule size and dry weights of beetles collected. Treatment had a significant effect on dry weight; mean dry weight decreased in Bt‐only treatments. Fisher's exact test of proportions of mating pairs of refuge and Bt insects indicated that mating was not random in 20% strip refuges and 5% seed blend treatments. We found high percentages of beetles that fed on Bt‐expressing plants as larvae, suggesting that mating between resistant beetles may not be rare even if random mating did occur.     

Differential mating success of virgin female katydidsRequena verticalis (Orthoptera: Tettigoniidae)

Male katydids (Orthoptera: Tettigoniidae) invest in offspring through nutrients provided in a large spermatophore. Previous research with Requena verticalishad shown that almost all of the investment of males mating with recently mated (4 days previously) females is in eggs fertilized by the female's previous mate. Thus males are predicted to discriminate against such females as mates. In experiments placing males with both a virgin and a female mated 4–5 days previously, virgin females obtained almost all matings. Although male discrimination of mates was noted in the experiments, there was no evidence that such discrimination was against nonvirgins in both this experiment and one in which a single virgin or mated female was placed with a male. Instead, the results suggest that the differential mating was a result of interfemale competition. The mating advantage held by virgin females over nonvirgins appeared to be lost once the latter had oviposited. Finally, there was no evidence from both single- and paired-female experiments that males preferred larger females as mates.     

Mortality of western corn rootworm larvae on MIR604 transgenic maize roots: field survivorship has no significant impact on survivorship of F1 progeny on MIR604

Mortality of western corn rootworm, Diabrotica virgifera virgifera LeConte, larvae due to MIR604 transgenic corn, Zea mays L., expressing the modified Cry3A (mCry3A) protein relative to survivorship on corn with the same genetic background without the gene (isoline corn) was evaluated at three Missouri sites in both 2005 and 2006. We made these comparisons by using wild-type western corn rootworm at three different egg densities (6,000, 3,000, and 1,500 eggs per m) so that the role of density-dependent mortality would be known. The mortality due to the mCry3A protein was 94.88% when averaged across all environments and both years. Fifty percent emergence of beetles was delayed approximately 5.5 d. Beetles were kept alive and their progeny evaluated on MIR604 and isoline corn in the greenhouse to determine whether survivorship on MIR604 in the field for one generation increased survivorship on MIR604 in the greenhouse in the subsequent generation. There was no significant difference in survivorship on MIR604 in greenhouse assays between larvae whose parents survived isoline and larvae whose parents survived MIR604 in the field the previous generation, indicating that many susceptible beetles survived MIR604 in the field the previous season along with any potentially resistant beetles. The data are discussed in terms of rootworm insect resistance management.     

Combinative stimulation inactivates sex pheromone production in the silkworm moth, Bombyx mori.

Mating results in a strong suppression of sex pheromone (bombykol) production in the female silkworm moth, Bombyx mori. The mechanical stimulation from the insertion of a penis, inflation of the bursa copulatrix (BC), or copulation with the sterile male whose penis was removed in order to prevent ejaculation (pr-male) induced only a partial decline in bombykol production. Artificial insemination stimulates oviposition of fertilized eggs as does normal mating. However, bombykol production did not decline in artificially inseminated females. When females were artificially inseminated before or after mating with pr-males, some females had a small amount of bombykol, similar to females mated with normal males, while other females had a large amount of bombykol similar to virgin females. The former usually laid fertilized eggs, while the latter laid only unfertilized eggs though semen filled their spermatophores and spermathecae. The mechanical stimulation caused by mating with a pr-male could be replaced by covering the abdominal tip with melted paraffin. Neither implantation of the BC obtained from mated females, nor injection of the spermatophore extract, into a female mated with a pr-male could inactivate bombykol production. Injection of hemolymph from a mated female into a virgin also failed to affect bombykol production These results indicate that a combination of both the tactile stimulation of the abdominal tip and the arrival of fertile spermatozoa in the vestibulum trigger a neural inactivation mechanism of bombykol production after mating.     

Juvenile hormone titre and egg production in Tenebrio molitor infected by Hymenolepis diminuta: effect of male and/or female infection,male age and mating

Infection of Tenebrio molitor with Hymenolepis diminuta induces curtailment of female fertility. We examined ovulation and oviposition, and associated titres of juvenile hormone (JH), in relation to parasitism and mating. Oviposition was significantly increased in infected mated and virgin beetles by days 6 and 9 post-emergence. Ovulation was not changed by infection; by the end of the 18-day experiment, the total number of laid eggs was not significantly altered. On day 6, JH levels were significantly higher in virgin infected insects, compared to non-infected controls (236+/-37.7 and 107+/-9.62 pg/g wet weight). Oviposition increased after mating, but total eggs ovulated remained the same. JH levels were higher in mated females on days 12 and 18 post-emergence, for infected and control insects. Previous studies suggested that male reproductive potential might rise following infection, because uninfected females lay more eggs when mated to infected males. We tested whether this caused an increase in female JH. Males were mated on days 5 or 12, when significant changes in their reproductive physiology begin to be observed, and are maximal, respectively. However, male age was of greater significance in promoting JH levels in females (p=0.001), than infection status of either partner (p=0.33).     

Chemosensory cues allow male Tenebrio molitor beetles to assess the reproductive status of potential mates

Males of many insect species, including beetles, choose their mates according to their reproductive status. However, the ways in which male beetles evaluate female reproductive status have received little attention. We tested the existence of male mate choice in the mealworm beetle, Tenebrio molitor, by observing mating and courtship behaviour of males given simultaneous access to pairs of females differing in their reproductive status: (1) mature versus immature; (2) virgin versus previously mated; (3) familiar (mated with the experimental male) versus unfamiliar (mated with a different male). Males courted and mated preferentially with mature and virgin females. To determine whether chemical cues played a role in these discriminations, we exposed males to filter paper squares bearing chemical cues from different types of females: (1) virgin versus mated; (2) mature versus immature. Males were significantly more attracted to those squares bearing chemical cues from virgin and mature females, suggesting that males can assess female reproductive status on the basis of chemical cues alone.     

Effects of different male remating intervals on the reproductive success of Choristoneura rosaceana males and females

The mass of the spermatophore transferred by a previously mated Choristoneura rosaceana male increases with time elapsed since the last mating but, even after 4 days, it never reaches the mass of the spermatophore of a virgin male. However, spermatophore mass is clearly not a good indicator of the male reproductive investment as the quantity of sperm in the second ejaculate of a previously mated male is the same as that of his first, if he is allowed a 2 (eupyrene sperm) to 3 day (apyrene sperm) recovery period. The interval between the first two matings had no influence on female fecundity or longevity but significantly affected fertility if the male had only 1 day to recover. The length of the post-copulatory refractory period was also shorter in females mated with previously mated males than in those mated with virgins, regardless of the male's remating interval. Furthermore, a significant variation in the eupyrene sperm content of the spermatophore transferred by virgin males had no influence on the length of the female refractory period. Globally, these results support the hypothesis that a factor, other than sperm numbers in the spermatheca, is responsible for maintaining the inhibition of pheromone production in this species.     

Polyandry in grain beetles, Tenebrio molitor, leads to greater reproductive success: material or genetic benefits?

Females that mate with more than one male may derive both materialand genetic benefits, and differentiating between the two benefitsis often difficult. We tested for both material and geneticeffects associated with multiple mating in the highly promiscuousyellow mealworm beetle, Tenebrio molitor. Females that matedfour times to the same male laid more eggs and produced morelarvae than females that mated only once. Whether copulationsoccurred on the same day or over several days, the result wasan immediate increase in the production of eggs by females.Some females were kept on a restricted diet to test whethernutrients in the spermatophore disproportionately benefittedfood-deprived females. Although females on poor diets producedfewer and smaller offspring, diet did not significantly affect the proportional benefit of mating treatment on female fecundity.By controlling for male mating history, we were able to separatethe effects of mating with different males from the effectsof receiving multiple spermatophores from the same male. Femalesthat mated with four different males achieved substantial gainsin numbers of eggs produced (32% increase) beyond those offemales that mated an identical number of times with the same male. We found no evidence that males allocate fewer sperm toprevious mates. Egg hatchability was unaffected by mating behavior,suggesting that genetic incompatibility at that stage is notresponsible for the low reproductive success of females matedwith a single male. These results suggest that females maydelay or reduce oviposition or may be incapable of achieving maximal fecundity until they have gained the material and/orgenetic benefits of mating with multiple males.     

Mating frequency and reproductive success in an income breeding moth,Mnesampela privata

The frequency of mating in insects is often an important determinant of female reproductive output and male sperm competition. In Lepidoptera that provide male nutrients to the female when mating, it is hypothesized that polyandry may be more prevalent. This is thought to be especially so among species described as income breeders; that is, in species who do not derive all their nutrients for reproductive output entirely from the resources obtained during the larval stage. We selected the geometrid moth, Mnesampela privata (Guenée) (Lepidoptera: Geometridae), to examine this hypothesis further. We found this species was best characterized as an income breeder with female weight on emergence positively correlated with total egg load but not with the number of eggs laid. Further, in accord with income breeders, females emerged with a partially developed egg load and lifetime fecundity was positively correlated with the number of oviposition days. However, in the laboratory we found that incidence of repeated matings or polyandry was rare. When moths were paired singly over their lifetime, only 4% of mated females multiple mated. When females were paired with three males concurrently, female mating success increased from 60 to 81% with multiple mating among mated females increasing to just 15%. Dissection of wild caught M. privata found that polyandry levels were also low with a maximum of 16.4% of females collected at any one time being multiple mated. In accord with theory, mating significantly increased the longevity of females, but not of males, suggesting that females acquire essential resources from male ejaculates. Despite this, multiple mated females showed a trend toward decreasing rather than increasing female reproductive output. Spermatophore size, measured on death of the female, was not correlated with male or female forewing length but was negatively correlated with the number of fertile eggs laid and female longevity. Smaller spermatophore width may be related to uptake of more nutrients by the female from a spermatophore. We discuss our findings in relation to income breeding and its relationship to polyandry in Lepidoptera.     

Western corn rootworm larval movement in SmartStax seed blend scenarios

Insect resistance management (IRM) can extend the lifetime of management options, but depends on extensive knowledge of the biology of the pest species involved for an optimal plan. Recently, the Environmental Protection Agency (EPA) registered seed blends refuge for two of the transgenic Bacillus thuringiensis (Bt) corn products targeting the western corn rootworm, Diabrotica virgifera virgifera LeConte. Larval movement between Bt and isoline plants can be detrimental to resistance management for high dose Bt products because the larger larvae can be more tolerant of the Bt toxins. We assessed movement of western corn rootworm larvae among four spatial arrangements of SmartStax corn (expressing both the Cry34/35Ab1 and Cry3Bb1 proteins) and isoline plants by infesting specific plants with wild type western corn rootworm eggs. Significantly fewer western corn rootworm larvae, on average, were recovered from infested SmartStax plants than infested isoline plants, and the SmartStax plants were significantly less damaged than corresponding isoline plants. However, when two infested isoline plants surrounded a SmartStax plant, a significant number of larvae moved onto the SmartStax plant late in the season. These larvae caused significant damage both years and produced significantly more beetles than any other plant configuration in the study (including isoline plants) in the first year of the study. This plant configuration would occur rarely in a 5% seed blend refuge and may produce beetles of a susceptible genotype because much of their initial larval development was on isoline plants. Results are discussed in terms of their potential effects on resistance management.     

Cost of reproduction in males of a satyrine butterfly Lethe diana

Adult butterflies feed largely on floral nectar and tree sap, both of which consist mainly of carbohydrates and include little nitrogen. They depend on the larval diet for nitrogenous resources. Consequently, there is a trade‐off between the reproductive and somatic nitrogenous investments of adults. Furthermore, male butterflies invest a considerable amount into spermatophores, containing nitrogen, which they give to their sexual partners. One way in which male butterflies could potentially replenish their spermatophores is by flight muscle histolysis, which may reduce locomotor ability and lifespan. In the present study, the effect of mating experience on nitrogen dynamics and the lifespan of males is investigated in the satyrine butterfly Lethe diana (Butler). Mated males do not have less thoracic nitrogen than virgin males, suggesting that mating experiences do not induce spermatophore recovery through flight muscle histolysis. Mated males possess less abdominal nitrogen than virgin males at death, indicating that they cannot recover the lost nitrogenous resource used for a single mating. Lifespan does not differ between mated and virgin males. Thus, reproduction and longevity are not fuelled necessarily by the same shared resource pools. Spermatophore mass increases as males get older. Considering that resources for producing spermatophores are limited, males may adjust the spermatophore mass strategically, depending on their chances of future reproduction.     

Oviposition behaviors in relation to rotation resistance in the western corn rootworm

Across a large area of the midwestern United States Corn Belt, the western corn rootworm beetle (Diabrotica virgifera virgifera LeConte, Coleoptera: Chrysomelidae) exhibits behavioral resistance to annual crop rotation. Resistant females exhibit increased locomotor activity and frequently lay eggs in soybean (Glycine max L.) fields, although they also lay eggs in fields of corn (Zea mays L.) and other locations. The goals of this study were (1) to determine whether there were any differences in ovipositional behavior and response to plant cues between individual rotation-resistant and wild-type females in the laboratory and (2) to examine the roles of, and interaction between, host volatiles, diet, and locomotor behavior as they related to oviposition. Because rootworm females lay eggs in the soil, we also examined the influence of host plant roots on behavior. In the first year of the study, rotation-resistant beetles were significantly more likely to lay eggs in the presence of soybean foliage and to feed on soybean leaf discs than wild-type females, but this difference was not observed in the second year. Oviposition by rotation-resistant females was increased in the presence of soybean roots, but soybean herbivory did not affect ovipositional choice. Conversely, ovipositional choice of wild-type females was not affected by the presence or identity of host plant roots encountered, and wild-type females consuming soybean foliage were more likely to lay eggs.     

Evaluation of Cuphea as a rotation crop for control of western corn rootworm (Coleoptera: Chrysomelidae)

The ability to prevent significant root feeding damage to corn, Zea mays L., by the western corn rootworm, Diabrotica virgifera virgifera LeConte, by crop rotation with soybean, Glycine max (L.) Merr., has been lost in portions of the Corn Belt because this pest has adapted to laying eggs in soybean fields. Cuphea spp. has been proposed as a new broadleaf crop that may provide an undesirable habitat for rootworm adults because of its sticky surface and therefore may reduce or prevent oviposition in these fields. A 4-yr study (1 yr to establish seven rotation programs followed by 3 yr of evaluation) was conducted to determine whether crop rotation with Cuphea would provide cultural control of corn rootworm. In support of Cuphea as a rotation crop, fewer beetles were captured by sticky traps in plots of Cuphea over the 4 yr of this study compared with traps in corn and soybean, suggesting that fewer eggs may be laid in plots planted to Cuphea. Also, corn grown after Cuphea was significantly taller during vegetative growth, had significantly lower root damage ratings for 2 of 3 yr, and had significantly higher yields for 2 of 3 yr compared with continuous corn plots. In contrast to these benefits, growing Cuphea did not prevent economic damage to subsequent corn crops as indicated by root damage ratings > 3.0 recorded for corn plants in plots rotated from Cuphea, and sticky trap catches that exceeded the threshold of five beetles trap(-1) day(-1). Beetle emergence from corn plots rotated from Cuphea was significantly lower, not different and significantly higher compared with beetle emergence from continuous corn plots for 2002, 2003 and 2004 growing seasons, respectively. A high number of beetles were captured by emergence cages in plots planted to Cuphea, indicating that rootworm larvae may be capable of completing larval development by feeding on roots of Cuphea, although peak emergence lagged approximately 4 wk behind peak emergence from corn. Based on these data, it is unlikely that crop rotation with Cuphea will provide consistent, economical, cultural control of corn rootworm.     

Multiple mating and lifetime reproductive output in female dobsonflies that receive nuptial gifts

In mating of the dobsonfly, Protohermes grandis (Thunberg), the male attaches the spermatophore externally to the female genitalia. The spermatophore includes a large gelatinous mass which the female detaches and feeds on after mating. While the female consumes this nuptial food gift, sperm is evacuated from the remaining portion of the spermatophore (sperm package) into her reproductive tract. Under laboratory conditions, mated females maintained receptivity throughout their lifetime, and they remated even on the day following copulation. A single insemination may supply enough sperm, as females mated only once deposited fertile eggs throughout life and, when dissected after death, all females had sperm in the spermatheca. There was a positive correlation between longevity and the number of matings. Lifetime fecundity also increased as mating multiplied. However, the size of eggs and hatchlings was not influenced by the number of matings. It seems that large spermatophore consumption by female P. grandis provides nutrients that increase fitness not in offspring quality, but in their quantity.     

Termination of sex pheromone production in mated females of the silkworm moth

A mating duration of more than 6 h was necessary to permanently terminate the production of the sex pheromone (bombykol) in the silkworm moth, Bombyx mori L. (Lepidoptera: Bombycidae), although the female formed a bursa copulatrix including a spermatophore and laid fertilized eggs even after mating for only 0.5 h. The 6-h mated female again produced bombykol if given an injection of synthetic pheromonotropic neuropeptide (PBAN), which is known to activate pheromone biosynthesis in a virgin female. Extracts of brain-suboesophageal ganglion (SG) complexes, which were removed from 6- and 24-h mated females, showed strong pheromonotropic activities. These results indicated that the pheromone gland of the mated female maintained its ability to biosynthesize bombykol; however, it could not produce pheromone due to a suppression of PBAN secretion from the SG. Furthermore, bombykol titers did not decrease after mating in females with a transected ventral nerve cord, even after the injection of a spermatophore extract, suggesting that the suppression of PBAN secretion was mediated by a neural signal and not by a substance in the spermatophore. The mated females accumulated (10E, 12Z)-10,12-hexadecadienoic acid, a precursor of bombykol biosynthesis, in their pheromone glands as did decapitated females. © 1996 Wiley-Liss, Inc.