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Journal of Insect Behavior, Vol. 17, No. 3, May 2004 (°


C 2004)

Exposure to Orange (Citrus sinensis L.)


Trees, Fruit, and Oil Enhances Mating
Success of Male Mediterranean Fruit Flies
(Ceratitis capitata [Wiedemann])
Todd Shelly,1−3 Charmian Dang,1 and Susan Kennelly2
Accepted: December 18, 2003; revised January 23, 2004

Previous laboratory tests revealed that exposure to oranges (Citrus sinensis


L.) increased the mating success of male Mediterranean fruit flies, Ceratitis
capitata (Wiedemann) (medfly). This advantage may have resulted from male
exposure to α-copaene (a sesquiterpene hydrocarbon and known male attrac-
tant) in the peel, as pure α-copaene has been shown to increase the mating
success of male medflies. Working with orange trees as well, we investigated
whether male exposure to nonfruiting trees, leaves (also known to contain
α-copaene albeit at a lower concentration than fruit), and fruit conferred a
mating advantage to wild-like males in field-cage tests. Males exposed to en-
tire nonfruiting trees or leaves had a mating advantage over control males
(exposed to a nonhost plant) in trials conducted 1 day but not 3 days after
exposure. Males exposed to orange fruits had higher mating success than con-
trol males (exposed to apples) in trials conducted 1 and 3 days after exposure.
Enhanced mating success was observed only when males were permitted to
contact the orange leaves and fruits; aroma alone did not affect male mating
success. In addition, we examined whether exposure to commercially available
orange oil, which also contains α-copaene, enhanced the mating performance
of wild-like and mass-reared sterile males. Heightened mating success was
observed in trials conducted 1 and 3 days after exposure for both types of

1USDA-APHIS, Waimanalo, Hawaii 96795.


2Center for Conservation Research and Training, University of Hawaii, Honolulu, Hawaii
96822.
3To whom correspondence should be addressed at USDA-APHIS, 41-650 Ahiki Street,
Waimanalo, Hawaii 96795. e-mail: todd.e.shelly@aphis.usda.gov.

303
0892-7553/04/0500-0303/0 °
C 2004 Plenum Publishing Corporation
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304 Shelly, Dang, and Kennelly

males, and in this case aroma alone had a positive effect on male mating suc-
cess. Future research should attempt to identify the behavioral, physiological,
or chemical mechanisms underlying the observed increases in male mating
success.
KEY WORDS: Mediterranean fruit fly; Ceratitis capitata; orange trees; mating success;
α-copaene.

INTRODUCTION

Host plants affect the mate searching strategy of male phytophagous insects
in two main ways. First, host plants may have a strong influence on male
spacing. In particular, males may concentrate their mate searching in the
vicinity of host plants if females aggregate there in search of food and/or
oviposition sites (Mangan, 1979; Greenfield, 1997). In addition, host plants
may affect male reproductive behavior via chemical interactions. Particular
plant-borne substances ingested by larvae (Lofstedt et al., 1989) or adults
(Krasnoff and Dussourd, 1989) may serve as precursors in the synthesis
of male sex pheromone. Exposure to host plant volatiles may also trigger
pheromone release in males (Jaffe et al., 1993) as well as increase female
responsiveness to male pheromones (Phillips et al., 1984).
Several recent studies have shown that plant-derived compounds have
an important effect on the mating behavior of male tephritids. For example,
males of the oriental fruit fly (Bactrocera dorsalis [Coquillett]) that feed on
pure methyl eugenol (Shelly and Dewire, 1994; Tan and Nishida, 1996) or
natural sources (flowers) of the compound (Shelly, 2000, 2002) have a pro-
nounced mating advantage over methyl eugenol-deprived males. Males that
consume methyl eugenol display increased signaling activity and produce a
more attractive pheromone (Shelly and Dewire, 1994), presumably result-
ing from the incorporation of certain metabolites of methyl eugenol in the
pheromone (Nishida et al., 1997). Growing evidence (Nishida et al., 1993;
Hee and Tan, 1998; Tan and Nishida, 2000) indicates that host plant sub-
stances influence male reproductive behavior in other species of Bactrocera
as well.
Similarly, work on the Mediterranean fruit fly (or medfly; Ceratitis cap-
itata [Wiedemann]), has revealed that feeding on wounded fruits of the or-
ange tree (Citrus sinensis L.) confers a mating advantage in laboratory tests
(Papadopoulos et al., 2001). The effect of fruit exposure was quite long-
lasting: males exposed to wounded oranges for 24 h had a mating advantage
over control males for at least 10 days after exposure. Likewise, Shelly and
Villalobos (2004) found that male medflies exposed to the bark and fruits of
guava trees (Psidium guajava L.) obtained significantly more matings than
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Mating Success of Male Medflies 305

(control) nonexposed males in tests conducted in field cages. Tests with both
orange and guava showed that males gained a mating advantage only when
they were permitted to contact the fruits (and, for guava, the bark) and that
exposure to fruit (or guava bark) aroma alone had no effect on male mating
success. In addition to natural substrates, exposure to particular essential
oils, namely, angelica seed oil and ginger root oil (Shelly, 2001), has been
shown to confer a mating advantage to male medflies. In contrast to the nat-
ural substrates, these oils conferred a mating advantage even when males
were prohibited from contacting them directly.
The chemical basis for these findings is unknown, but evidence suggests
that α-copaene, a plant-borne sesquiterpene hydrocarbon and known attrac-
tant of male medflies (Flath, 1994a,b), is involved. The substances shown to
confer a mating advantage to male medflies all contain this chemical (or-
ange fruit [Teranishi et al., 1987], guava fruit [Oliveros-Belardo et al., 1986],
guava bark [F. Webster, personal communication], angelica seed and gin-
ger root oils [Takeoka et al., 1990]). In addition, male medflies exposed to
pure α-copaene had a mating advantage over control, nonexposed males
(Shelly, 2001). (Although α-copaene is clearly implicated, it should be rec-
ognized that many related terpenoids co-occur with α-copaene and may
also affect the behavior of male medflies either independently or in com-
bination with α-copaene [Flath, 1994a,b)]. Consequently, any reference to
α-copaene is perhaps best considered to include not only that compound,
but other sesquiterpenes that may also influence male medflies.) How α-
copaene enhances male mating performance is not known, although tests
with ginger root oil showed that exposed males signaled more frequently
(Shelly, 2001) and mounted females more quickly (i.e., exhibited shorter
courtship [Briceno et al., unpublished data]) than control males. Exposure
to ginger root oil does not appear to affect the attractiveness of the male sex
pheromone (Shelly, 2001).
The present study expanded upon that of Papadopoulos et al. (2001)
and focused on two main objectives regarding the interaction between or-
ange trees and male medflies. First, we investigated whether male exposure
to nonfruiting trees, leaves, and fruit conferred a mating advantage to wild-
like males (see below) in field-cage tests. Nishida et al. (2000) reported that
α-copaene occurs in the leaves of various citrus plants, including orange,
and consequently we investigated whether exposure to entire, nonfruiting
trees or leaves only increase male mating success. Second, we examined
whether exposure to commercially available orange oil, which also contains
the attractant α-copaene, enhanced the mating performance of both wild-
like and mass-reared, sterile males. The release of sterile male medflies is an
important means of suppressing infestations of this notorious agricultural
pest (Hendrichs et al., 2002), and exposure to ginger root oil has been shown
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306 Shelly, Dang, and Kennelly

to increase the mating performance of sterile males relative to wild males


(Shelly and McInnis, 2001). Here, we determine whether prerelease expo-
sure to orange oil similarly increases the mating competitiveness of sterile
males.

METHODS

Study Flies. Wild-like flies were derived from a laboratory colony started
with 200–300 adults reared from coffee (Coffea arabica L.) collected near
Haleiwa, Oahu. The colony was maintained in a screen cage and provided
with ample food (sugar and yeast hydrolysate at a 3:1 volumetric ratio),
water, and oviposition substrate (perforated plastic vials containing small
sponges soaked in lemon juice). Eggs were placed on standard larval medium
(Tanaka et al., 1969) in plastic containers over vermiculite for pupation.
Adults used in the mating experiments were separated by sex within 24 h of
emergence, well before reaching sexual maturity at 5–7 days of age. When
used in this study, the wild-like flies were six to eight generations removed
from the wild.
Mass-reared males were from the Maui-Med strain produced by the
USDA-APHIS Hawaii Fruit Fly Rearing Facility, Waimanalo, HI, since
1996 (i.e., mass-reared for approximately 60 generations prior to this study).
Pupae of this mass-reared strain were exposed in air to 15 krad of gamma
irradiation from a 137 Cs source 2 days before eclosion and then delivered
to the laboratory. Males of this strain were collected within 12 h of emer-
gence (sexual maturity is attained at 2–3 days of age [D. O. McInnis, personal
communication]). Both wild-like and mass-reared adults were held in plas-
tic buckets covered with nylon screening (5-liter volume, 100–150 flies per
bucket) with ample food and water at 22–25◦ C and 65–85% RH with a pho-
toperiod of 12:12 (L:D) from natural and artificial light.
Male Exposure to Orange Trees, Leaves, and Fruit. We ran three ex-
periments that involved wild-like flies exclusively. In the first, we exam-
ined whether exposure to nonfruiting orange trees influenced male mat-
ing success. We released 150 (treated) males between 0700 and 0730 into
nylon-mesh field cages (3-m diameter, 2.5 m high) that contained two pot-
ted, nonfruiting orange trees (approximately 2 m tall). The field cages were
located at the Agricultural Experiment Station of the University of Hawaii,
Waimanalo. Five trees, presented in different combinations, were used for
exposing males. Males were collected 4 h later, returned to the laboratory
(where food and water were provided), and then tested 1 or 3 days later.
Control males were treated in the same manner except that they were re-
leased in a field cage containing a nonhost tree (fiddlewood, Citharexylum
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Mating Success of Male Medflies 307

spinosum L.). A single fiddlewood tree was used for all control males. Air
temperature (1 m aboveground in the shade) ranged from 23 to 32◦ C during
the exposure period. When released for exposure, treated and control males
were 8–11 days old.
In the second experiment, treated males were tested after exposure to
fresh leaves in the laboratory. Leaves were removed from trees near the
laboratory and immediately placed in screen cages (30-cm cubes). Forty
leaves were used per cage, and these were placed on chicken-wire screen
raised above the cage floor, allowing the males access to both top and bottom
leaf surfaces. We then placed 60 males in individual cages at 0800, removed
them 3 h later, and held them (with food and water) until tested 1 or 3 days
later. Control males were handled in the same manner except that they
were exposed to leaves (40 leaves/cage) from a fiddlewood tree (the leaves
used were of similar size to orange leaves). When exposed to the leaves,
treated and control males were 8–13 days old. Treated and control males
were exposed to leaves in separate rooms to avoid inadvertent exposure
of orange aroma to the control males (both rooms were isolated from the
colony as well). In an additional test, we covered the orange leaves with
nylon screening to prevent the males from contacting them. Males treated
in this manner were tested 1 day later.
Third, treated males were exposed to fruit in the laboratory. Sixty males
were placed in screen cages (30 cm cubes) at 0800 with two Valenica oranges,
collected 3 h later, and kept in the laboratory (with food and water) until
tested 1 or 3 days later. As with leaf exposure, we modified this protocol
by placing the two oranges in screen containers that allowed males to de-
tect the fruit odor but prevented them from making direct contact with the
fruit. Males treated in this manner were tested 1 day later. Control males
were handled in the same manner except that they were presented with two
Granny Smith apples (Malus sylvestris Mill.) instead of oranges. Fruit were
purchased in a supermarket and then rinsed in water and dried prior to use.
Also, before exposure, we made five shallow cuts (2–4 cm long) into the peel
of all fruit using a scalpel. When exposed to fruit, treated and control males
were 8–10 days old. As with the leaf exposure, treated and control males
were exposed to fruit in separate rooms.
Male Exposure to Orange Oil. We ran three experiments involving male
exposure to orange oil (Oil Orange California Type C.P. FCC; Product code,
15061; Citrus and Allied Essences, Lake Success, NY), containing α-copaene
0.2% (concentration [S. Young, personal communication]). A standard ex-
posure protocol was followed in all experiments. We applied 20 µl of the
oil to a small disk of filter paper using a microcapillary pipette. The disk
was placed on the bottom of a transparent plastic drinking cup (400-ml
volume), 25 males were immediately placed in the cup using an aspirator,
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308 Shelly, Dang, and Kennelly

and the cup was covered with nylon screening. Exposure started at 0700
and continued until 1100. The behavior of the males was not monitored
systematically during exposure periods, but in frequent checks males were
never observed touching the paper disc (the same observation was made
for ginger root oil [Shelly, 2001]). In a given experiment, control males
were simply nonexposed males of an age similar to that of treated (ex-
posed) males. Exposure to the oil was always conducted in an isolated
room to prevent inadvertent exposure to control males (and the colony
as a whole).
In the first experiment, we compared the mating success of treated
and control wild-like males. Treated males were exposed at 8–10 days of
age using the standard method and tested 1 or 5 days later, respectively.
In the second experiment, treated and control mass-reared males competed
separately (i.e., in separate field cages) against wild-like males. Treated mass-
reared males were exposed at 4–5 days of age to orange oil using the standard
protocol and tested 1 or 5 days later, respectively. Wild-like males (9–14 days
old when tested) were not exposed to orange oil in this experiment. In the
third experiment, treated and control mass-reared males competed directly
against one another (i.e., no wild-like males were used). Treated males were
exposed to orange oil using the standard protocol or, as with the orange
leaves and fruits, were exposed to the aroma only. In both cases, treated
males were 4–5 days old when exposed and were tested 1 day later. To
present direct contact with the oil, we placed the oil-laden disks in small
containers (covered with nylon mesh screening) that were then introduced
into the plastic cups.
Mating Trials. Mating trials were conducted in Waimanalo, Oahu, at the
same University of Hawaii facility noted above. In all experiments (with one
exception), groups of 100 treated and 100 control males of the same strain
(i.e., wild-like or mass-reared) and 100 wild-like females were released be-
tween 0700 and 0730 in field cages that contained a single rooted guava
tree. (In the exception, 100 mass-reared and 100 wild-like males were re-
leased along with 100 wild-like females in one experiment testing the effect
of orange oil exposure. In this case, mass-reared males were either treated
or control, and wild-like males were never treated). For a given test, we
marked either treated or control males (or mass-reared or wild-like males
for the aforementioned exception), alternating the marked group between
successive tests. Males were marked 1 day before testing by cooling them
for several minutes and placing a dot of enamel paint on the thorax. This
procedure had no obvious adverse effects, and males resumed normal activ-
ities within minutes of handling. The cages were monitored for 4 h, mating
pairs were collected in vials, and the males were identified. Wild-like females
(8–14 days old) were used in all tests. All flies were used only once.
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Mating Success of Male Medflies 309

Statistical Analyses. Pairwise comparisons were made using the t-test.


Assumptions of normality (tested using the Kolmogorov–Smirnov distri-
bution with the Lillefors correction) and homoscedasticity (tested using
Levene’s median test) were met in all cases. Where comparisons involved
proportions, values were arcsine transformed. Statistical tests were per-
formed using SigmaStat (Version 2.0).

RESULTS

Orange Trees, Leaves, and Fruit. Wild-like males exposed to orange


trees obtained significantly more matings than (control) wild-like males ex-
posed to a nonhost tree in tests conducted 1 day following exposure (Table I).
However, exposure to orange trees conferred no mating advantage in tests
conducted 3 days following exposure. Similar results were obtained when
wild-like males were exposed to leaves in laboratory cages: exposure to
orange leaves (with contact possible) conferred a mating advantage 1 day
but not 3 days after exposure (Table I). When denied contact with the or-
ange leaves, however, no difference in mating success was detected between
treated and control males. Wild-like males exposed to orange fruit achieved
significantly more matings than (control) wild-like males exposed to apples,
and in this instance a mating advantage was evident both 1 and 3 days after

Table I. Mating Success of Treated Versus Control Wild-like Males, Where Treated Males
Were Exposed to Nonfruiting Orange Trees in Field Cages (Control Males Were Exposed to a
Fiddlewood Tree) or Orange Leaves and Fruit in the Laboratory (Control Males Were Exposed
to Fiddlewood Leaves and Apples, Respectively)

Matings per replicate


Exposure Postexposure
Experiment treated males interval (days)a Treated Control t

1 Trees 1 33.0 (8.3) 18.8 (6.2) 4.1∗∗


3 24.6 (5.0) 25.9(7.2) 0.4 (NS)
2 Leaves 1 29.9 (7.8) 13.5 (4.3) 4.9∗∗∗
3 19.1 (4.7) 15.9 (4.6) 1.0 (NS)
1(hc) 17.8 (6.9) 18.9 (8.8) 0.4 (NS)
2 Fruit 1 26.9 (7.7) 10.4 (5.3) 5.0∗∗∗
3 23.2 (6.7) 11.9 (3.8) 4.2∗∗∗
1(nc) 23.5 (7.7) 18.0 (9.9) 1.2 (NS)

Note. Values represent means (±1 SD) based on nine replicates for tree exposure and eight
replicates for both leaf and fruit exposure. See text for details of exposure protocols.
a For treated males, days elapsed between exposure and testing; control males were approxi-
mately the same age as treated males in all experiments. (HC) No contact of orange leaves or
fruit was possible.
∗∗ P < 0.001; ∗∗∗ P < 0.001. NS, not significant, at P = 0.05.
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310 Shelly, Dang, and Kennelly

exposure (Table I). However, treated males accounted, on average, for a


slightly higher proportion of the total matings 1 day after exposure than
3 days after exposure (75 versus 66%, respectively; t = 2.4, P < 0.05).
The advantage conferred by fruit exposure apparently derived from physi-
cal contact with the oranges, because exposure to fruit odor alone did not
enhance the mating success of treated males (Table I).
Orange Oil. In competition between wild-like males (Table II; Exper-
iments 1a and 1b), individuals exposed to orange oil achieved significantly
more matings than nonexposed individuals in tests performed 1 or 5 days
after exposure of the treated males.
The mating success of mass-reared males relative to wild-like males de-
pended strongly on whether the mass-reared males were exposed to orange
oil (Table II; Experiments 2a and 2b). Mass-reared males exposed to orange
oil had a mating advantage whether tested 1 or 5 days after exposure, ac-
counting for 70 and 65% of the total matings, respectively (t = 0.3, P > 0.05).
In contrast, when neither group was exposed to orange oil, (control) wild-
like males outcompeted the (control) mass-reared males associated with
both postexposure intervals.

Table II. Mating Success of Wild-like and Mass-Reared Males, Where Treated Males Were
Exposed to Orange Oil (Control Males Were Not Exposed)

Postexposure
Experiment Male type interval (days)a Mating per replicate t

1a Wild-like, treated 1 35.7 (10.4) 4.7∗∗∗


Wild-like, control 15.4 (6.5)
1b Wild-like, treated 5 30.2 (6.9) 4.1∗∗∗
Wild-like, control 18.6 (7.3)
2a Mass-reared, treated 1 27.2 (9.4) 4.8∗∗∗
Wild-like, control 11.5 (6.6)
Mass-reared, control 14.5 (7.6) 3.9∗∗
Wild-like, control 35.1 (11.2)
2b Mass-reared, treated 5 30.0 (10.2) 3.8∗∗
Wild-like, control 16.0 (7.4)
Mass-reared, control 11.6 (6.6) 4.2∗∗∗
Wild-like, control 24.9 (8.8)
3a Mass-reared, treated 1 33.6 (8.1) 4.9∗∗∗
Mass-reared, control 14.0 (8.0)
3b Mass-reared, treated 1 (nc) 32.8 (7.2) 4.3∗∗∗
Mass-reared, control 16.7 (7.7)

Note. Values represent means (±1 SD) based on eight replicates for all experiments. See text
for details of exposure protocol.
a For treated males, days elapsed between exposure and testing; control males were approx-
imately the same age as treated males in all experiments. (nc) No contact with orange oil-
containing disk possible.
∗∗ P < 0.001; ∗∗∗ P < 0.001.
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Mating Success of Male Medflies 311

Consistent with the preceding experiment, treated mass-reared males


had a pronounced mating advantage over control mass-reared males in direct
competition, and this pattern was evident regardless of whether the treated
males were able to contact the oil-containing disk (Table II; Experiment 3).
On average, the proportion of matings obtained by treated males able to
contact the disk was similar to that for treated males denied contact (70 and
66%, respectively; t = 0.2, P > 0.05).

DISCUSSION

Consistent with earlier data (Papadopolous et al., 2001), the present


study shows that (1) exposure to orange fruit conferred a mating advan-
tage to male Mediterranean fruit flies, (2) this advantage lasted several days
after exposure, and (3) direct contact with fruit was required for mating
enhancement, i.e., exposure to fruit odor alone had no effect. Similar ef-
fects were obtained following male exposure to guava fruit as well (Shelly
and Villalobos, 2004). The present study also demonstrated that male ex-
posure to entire nonfruiting orange trees and leaves alone increased male
mating success. In contrast to fruit exposure, however, the positive effect on
male mating was short-lived, and there was no significant difference between
control and treated males in tests conducted 3 days after treated males were
exposed to orange trees or leaves. This difference between fruit and leaf
exposure may reflect a difference in the concentration of α-copaene in these
structures: orange peels contain approximately 0.7 µg/g of α-copaene (R.
Nishida, personal communication), compared to only 0.3–0.4 µg/g leaf for
orange leaves (Nishida et al., 2000; comparable data are not available for
guava).
The present findings for orange (along with those for guava [Shelly and
Villalobos, 2004]) suggest that chemical interactions between males and host
plants may influence the distribution of male mating aggregations (leks) in
the environment. Although field data on spacing of male and female medflies
are scant, evidence (summarized by Field et al., 2002) indicates that males
tend to cluster at sites of high female traffic (or hotspots [Bradbury and
Gibson, 1983]). According to this idea, medfly leks generally from on par-
ticular host trees, because females are attracted there for oviposition. Thus,
although males do not control access to fruit, they perch nearby to increase
encounter rates with receptive females requiring sperm prior to oviposition.
While female response to oviposition sites undoubtedly affects male distri-
bution, the present study suggests that chemical interactions between males
and host plants may also influence male spacing independently of female
movement.
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312 Shelly, Dang, and Kennelly

Moreover, because the nature of these male-plant interactions varies


among different host plant species, there may be corresponding variation in
the degree of congruence between male and female responses to a given host
plant. In the case of orange, for example, the presence of fruits is likely to
attract both sexes strongly, because males gain a longer-lasting mating benefit
from fruit than leaves, and females require fruit for oviposition. In the case
of guava, however, males gain equal mating benefits through contact with
ripe fruit or patches of bark rich in α-copaene (Shelly and Villalobos, 2004).
Thus, while fruiting guava trees might attract both sexes, nonfruiting trees
may still be attractive to males (if these contains high levels of α-copaene)
but not, per se, to females (i.e., independent of male presence). Because of
the complex interplay between host plant chemistry and medfly behavior,
experimental studies that manipulate the distribution of oviposition sites and
α-copaene sources independently may be required to evaluate the relative
importance of these two resources on lek distribution.
Like ginger root oil (Shelly, 2001), exposure to orange oil greatly in-
creased the mating success of male medflies. In fact, orange oil reversed the
outcome of mating competition between wild-like and mass-reared, sterile
males. Without exposure to orange oil, mass-reared males achieved about
one-third of the total matings, whereas after exposure they obtained approx-
imately two-thirds of all matings. Also, increased mating success was evident
even when males were prohibited from contacting the orange oil (the same
result was found for ginger root oil [Shelly, 2001]). These findings suggest
that, like ginger root oil (Shelly and McInnis, 2001), orange oil could po-
tentially be used to increase the mating competitiveness of sterile males in
control programs against the medfly. To further investigate this possibility,
the effectiveness of orange oil should be examined at large-scale exposure.
Specifically, sterile male release programs against medfly often hold newly
emerged flies for 3–4 days in large holding boxes (so-called PARC boxes),
each of which holds approximately 40,000 males. The entire box could be ex-
posed to orange oil (by placing oil-laden filter paper on a screened opening
on the top of the box), and the mating success of these treated males could
be compared to control males from unexposed boxes.
Preliminary evidence suggests that, like their wild counterparts, mass-
reared males of C. capitata gain a mating advantage through contact with
guava and orange fruits. Thus, sterile males may receive a competitive boost
independent of any prerelease exposure regime. Even so, of course, pre-
release exposure to orange (or ginger root) oil eliminates the “need” for
sterile males to locate chemical sources in the environment (thereby elim-
inating time and energy costs associated with searching) and guarantees
that sterile males benefit fully from exposure to a performance-enhancing
oil.
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Mating Success of Male Medflies 313

Future work should focus on identifying the mechanism underlying in-


creased male mating success. Field (Shelly, 2001) and laboratory
(Papadopoulous and Shelly, unpublished data) experiments using ginger
root oil revealed no difference in long-range female attraction to exposed
versus nonexposed males. A chemical perspective also argues against a
role for α-copaene (or metabolites) in pheromone synthesis, because no
compound similar in structure to α-copaene has been recorded in male-
produced volatiles (Millar, 1995). One possibility is that medfly males alter
the pheromonal composition during courtship (i.e., following female arrival)
and that α-copaene plays a role in this short-range communication (W. G.
Eberhard, personal communication). Data (Briceno et al., unpublished data)
showing quick female acceptance of ginger root oil–exposed males are con-
sistent with this idea. Comparing the mating success of exposed and nonex-
posed males when courting normal females versus females whose ability to
sense pheromones has been altered would offer rigorous evaluation of this
hypothesis.

ACKNOWLEDGMENTS

We thank Roger Coralis for permission to conduct fieldwork at the agri-


cultural station, Erik Rutka and Mindy Teruya for assistance in the labora-
tory and field, James Duke, Ritsuo Nishida, Guadalupe Rojas, Fran Webster,
and Silvia Young for information on α-copaene, and Bill Eberhard, Don
McInnis, and Nikos Papadopoulos for helpful discussions. This research was
supported in part by a grant from the Binational Agricultural Research and
Development (BARD Project No. US-3256-01) to B. Yuval and T.E.S.

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