Field evaluation of attractive lures for the fruit fly Bactrocera minax (diptera: tephritidae) and their potential use in spot sprays in Hubei province (China)

Field Evaluation of Attractive Lures for the Fruit Fly Bactrocera
minax (Diptera: Tephritidae) and Their Potential use in Spot
Sprays in Hubei Province (China)
Author(s): Xiao-Wei Zhou, Chang-Ying Niu, Peng Han, and Nicolas Desneux
Source: Journal of Economic Entomology, 105(4):1277-1284.
Published By: Entomological Society of America
/>URL: />
BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the
biological, ecological, and environmental sciences. BioOne provides a sustainable online
platform for over 170 journals and books published by nonprofit societies, associations,
museums, institutions, and presses.
Your use of this PDF, the BioOne Web site, and all posted and associated content indicates
your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use.
Usage of BioOne content is strictly limited to personal, educational, and non-commercial
use. Commercial inquiries or rights and permissions requests should be directed to the
individual publisher as copyright holder.

BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers,
academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research.


FIELD AND FORAGE CROPS

Field Evaluation of Attractive Lures for the Fruit Fly Bactrocera minax
(Diptera: Tephritidae) and Their Potential Use in Spot Sprays in
Hubei Province (China)
XIAO-WEI ZHOU,1 CHANG-YING NIU,1,2 PENG HAN,1,3

AND

NICOLAS DESNEUX3

J. Econ. Entomol. 105(4): 1277Ð1284 (2012); DOI: />
ABSTRACT The Chinese citrus fruit ßy, Bactrocera minax (Enderlein) is a univoltine Tephritidae
pest that infests Citrus species. Field trials were conducted in 2010 to determine the potential use of
a lure based on enzymatical-hydrolyzed beer yeast as liquid bait (hereafter named H-protein bait)
for B. minax in the Hubei province, China. In a citrus orchard, we compared the attractiveness among
aqueous solutions of H-protein bait, GF-120 fruit ßy bait, sugar-vinegar-wine mixture, torula yeast, and
Jufeng attractant when used in traps and in spot sprays, that is, lures used in combination with the
insecticide trichlorphon. The H-protein bait was the most attractive lure in traps, ensnaring signiÞcantly more adults than sugar-vinegar-wine mixture, torula yeast, and Jufeng attractant, in decreasing
efÞciency order. In spot sprays those with H-protein bait killed signiÞcantly more female and male
ßies within 40 min than those with sugar-vinegar-wine mixture, GF-120, Jufeng attractant, and the
control. In addition, the total number of ßies killed by H-protein bait during the spot spray duration
was higher than other treatments. Our results demonstrated that the H-protein bait may be a useful
tool in citrus orchards in China to monitor B. minax populations as well as to manage this pest when
used in spot sprays.
KEY WORDS hydrolyzed-protein bait, GF-120, phenology, citrus orchard, monitoring

The Chinese citrus fruit ßy, Bactrocera minax (Enderlein) (Diptera: Tephritidae) has been a serious pest on
citrus fruits in China, India (West Bengal and Sikkim),
and Bhutan for more than half a century (Drew 1979,
Wang and Luo 1995, Dorji et al. 2006). Among ßies of
the Dacinae subfamily (Tephritidae family) B. minax
has the particularity of being a univoltine species
(Dorji et al. 2006), that is, one generation per year.
The host range of B. minax is restricted to the Citrus
species and adult emergence usually occurs from late
April to early May, with adults present in the Þeld until
August in China. Adults lay eggs on immature citrus
fruits from mid-June to July. The phenology varies
among infested regions and appears to vary according

to local temperatures (Wang and Luo 1995). In the
Hubei province, B. minax is considered one of the
major fruit ßy pests on citrus trees, causing considerable economic damage by making the fruits improper
for merchandizing (Yang et al. 1994). Therefore, effective Integrated Pest Management (IPM) programs
are required for areas where the citrus production is
a major economic issue.
1 Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science & Technology, Huazhong Agricultural University, Wuhan 430070, China.
2 Corresponding author, e-mail:
3 French National Institute for Agricultural Research (INRA), 400
route des chappes, 06903 Sophia-Antipolis, France.

Traps, attractive lures, and mass trapping techniques are common strategies for management of fruit
ßies (Hendrichs et al. 1995, Stonehouse et al. 2002,
Vargas et al. 2009). For example, McPhail traps baited
with a fermenting mixture of citrus juice and brown
sugar were successfully used to trap fruit ßies in early
studies in Florida (Newell 1936). In addition, an approach combining olfactory attractants and baits
sprayed with insecticides, that is, lures ϩ insecticides
has been used to combine detection and monitoring in
the management of fruit ßies. Protein baits sprayed
with insecticides were Þrst used for Caribbean fruit ßy
control (Steiner 1952). Since then, protein bait sprays
have become a major method of suppressing or eradicating fruit ßy populations in many parts of the world.
Female tephritids require a protein meal for ovarian
development and egg production (Christenson and
Foote 1960, Mangan 2003, Perez-Staples et al. 2007)
and protein sources, such as bacteria in bird feces, are
thought to be scarce in natural conditions (Drew et al.
1983). Therefore, protein sources are highly attractive
to tephritid females and thus protein bait sprays have

been successfully used to manage tephritid species
(Yee and Chapman 2005; Mangan et al. 2006; Vargas
and Prokopy 2006; McQuate 2009; Pin˜ ero et al.
2009a,b). Two commonly used protein baits for fruit
ßy detection, monitoring, management, and trapping
are the GF-120 Fruit Fly Bait (Dow AgroSciences,
Indianapolis, IN) and torula yeast (ERA International,

0022-0493/12/1277Ð1284$04.00/0 ᭧ 2012 Entomological Society of America


1278

JOURNAL OF ECONOMIC ENTOMOLOGY

Freeport, NY) (Burns et al. 2001; Vargas et al. 2002;
Prokopy et al. 2003; Stark et al. 2004; Mangan et al.
2006; McQuate 2009; Pin˜ ero et al. 2009a,b). To our
knowledge, potential use of protein baits to manage B.
minax in China has not been documented. In most
places where B. minax is present in this country, farmers frequently rely on spraying sugar-vinegar-wine
mixture with insecticides to manage the pest (Wang
and Luo 1995). Though most fruit ßies can be monitored by traps baited with sexual pheromone and parapheromones, B. minax is not known to be attracted to
any male lures (Drew et al. 2006). How the GF-120
and torula yeast can be effective against B. minax in
China has not yet been documented. By contrast, the
Jufeng attractant is registered and commercially available as lure for B. minax in China though its efÞcacy
has never been clearly demonstrated. In our laboratory, a new protein bait has recently been developed
to control B. minax. It is based on an enzymatically
hydrolyzed protein produced by the industrial processing of beer yeast. This hydrolyzed protein bait

(hereafter named H-protein bait) also includes feeding stimulants, orange juice and brown sugar, as well
as a chemical attractant ammonium acetate.
In 2010, in Þeld conditions we assessed the effectiveness of the H-protein bait and several other commercially available lures for potential monitoring and
management of B. minax. In a citrus orchard, two
experiments were performed: 1) we compared the
efÞcacy of the H-protein bait, torula yeast, sugar-vinegar-wine (SVW) mixture, and Jufeng as attractant for
B. minax, and 2) we studied how various lures (Hprotein bait, SVW mixture, GF-120, and Jufeng as an
attractant) are effective against B. minax when used in
spot sprays, that is, when combined with an insecticide. As a hand sprayer was needed to create the spot
sprays, torula yeast that did not completely dissolve in
water (it is provided as pellets) could not be used in
this experiment. GF-120 was used only in the second
experiment as a positive control (it is commonly used
for spot sprays in various crops or orchards).
Materials and Methods
Field Site. The study was conducted from early May
to early August 2010, during the fruiting season, in a 0.4
ha citrus orchard in Yichang, in the Hubei province,
China. The orchard was composed of mainly mandarin
(Citrus reticulata), navel orange (Citrus sinensis), and
pomelo (Citrus maxima) trees. Density of trees in the
orchard was Ϸ1,000 trees per ha. No management
practice for B. minax or other pest control was carried
out in the orchard during the experimental periods,
except for routine management such as grass-mowing
and pruning. The average daily temperature and relative humidity during the experiments were 27.2 Ϯ
0.3ЊC, 67.5 Ϯ 4.3%.
Lures. The lure treatments were prepared using the
following formulations: 1) H-protein bait: 20% vol:vol
solution in water; 2) SVW mixture: 3% sugar solution

with vinegar and wine mixture; 3) torula yeast: two
pellets per 300 ml of water; 4) GF-120 fruit ßy bait: 1:3

Vol. 105, no. 4

(vol:vol) solution at the recommended application
rate; and 5) Jufeng attractant: 25 g of lure in 300 ml
water as recommended. The pH of lures was checked
after preparation using a pH meter (PB-10, Sartorius,
Germany).
Experiment 1: Assessment of Attractiveness of the
Lures in Citrus Orchard. The experiment was set up
on 4 May 2010 and trapping was carried out periodically, once a week, until 10 August 2010. Four types of
lures were tested for attractiveness in traps: H-protein
bait, SVW mixture, torula yeast, and Jufeng attractant.
The traps were made of modiÞed plastic water bottles
(height: 18 cm, diameter: 9 cm, a 4 ϫ 3 cm window 8
cm from the bottom of the bottle). This design ensnares adult ßies which are unable to escape and
drown in water or die because of starvation. The traps
were baited with 250 ml of aqueous solution and attached to citrus tree branches 1.5 m in height. The
traps were located randomly within the orchard with
a space of 15 m separating the traps. The trials were set
up in a randomized complete-block design using nine
replicates (traps) per lure type, that is, 4 ϫ 9 ϭ 36 traps
were used every week. Once a week, when traps were
checked and ßies were removed, the traps were thereafter washed with water and lures renewed. At the
same time, each trap was moved to a new position
within the experimental site. The number of male and
female B. minax captured was recorded and the ßies
were placed in 75% ethanol. The females were subsequently dissected to estimate their sexual maturity

under a stereomicroscope (Nikon Inc., SMC-10, Japan) and placed into three categories: immature (ovaries small and no eggs present), semimature (ovaries
with eggs developing but not mature), and mature
(ovaries with mature eggs present). This classiÞcation
was based on preliminary laboratory experiments
(C.Y.N., unpublished data).
The number of male and female ßies trapped per
week was analyzed using a generalized linear model
(Proc Genmod; SAS Institute 1999) with ÔLure,Õ ÔSex,Õ
ÔDateÕ as factors. Potential interactions among these
factors were also tested. In addition, we tested for a
potential relationship between the numbers of B. minax female ßies ensnared in H-protein bait traps and
the sexual maturity of these ßies, using regression
analyses. This analysis was carried out to assess the
attractiveness of the H-protein bait in relation to the
sexual maturity of B. minax females.
Experiment 2: Assessment of Effectiveness of Lures
When Used in Spot Sprays. Spot spray tests were
conducted from 4 June to 14 June in 2010 in the citrus
orchard. The test was designed to study the speed and
the overall effectiveness of four different lures when
used in spot sprays under Þeld conditions. Five lureinsecticide solutions, that is, spot sprays, were prepared: the H-protein bait, the SVW mixture, the
Jufeng attractant, the GF-120 (positive control), and
water (control), and all solutions were supplemented
with the Trichlorphon 90% WP insecticide (Dacheng
Pesticide Co., Ltd., Shandong, China) at the rate of
0.12 g per 100 ml of solution. For each replicate (i.e.,
spot spray), 150 ml solutions (with lures and insecti-


August 2012

ZHOU ET AL.: ATTRACTIVE LURES FOR THE FRUIT FLY Bactrocera minax

Table 1. Statistics from the generalized linear model used to
analyze the numbers of flies
Source of variationa
A: Lure attractiveness
Lure
Sex
Date
Lure ϫ Date
Lure ϫ Sex
Date ϫ Sex
B: Spot spraysÑimmediate effect
Lure
Sex
Lure ϫ Sex
C: Spot spraysÑtotal effect
Lure
Sex
Lure ϫ Sex

df

␹2

P value

3
1

10
30
3
10

143.09
0.09
283.24
654.01
0.91
16.34

Ͻ0.001
0.761
Ͻ0.001
Ͻ0.001
0.823
0.090

4
1
4

63.05
3.30
5.35

Ͻ0.001
0.069
0.253

4
1
4

46.57
10.71
6.21

Ͻ0.001
0.001
0.184

a
(A) trapped during the course of the exp, (B) found dead on the
spot sprays during the Þrst 40 min after initial spray application (i.e.
immediate effect), and (C) killed on the spot sprays until no dead ßies
were longer observed on the spots (i.e. persistent effect) among the
various lures tested (Lure factor), as function of ßies sex (Sex factor)
and also as function of date of sampling (Date factor) in case of the
Lure attractiveness experiment.

cide) were applied to foliage of two citrus trees using
hand-held sprayers (500 ml in capacity) (Farm and
Garden Machinery Sales Co., Jinhua, China). The tests
were carried out at 10:30 a.m., that is, when B. minax
shows its highest diurnal activity (C.Y.N. and X.W.Z.,
unpublished data). Three replicates were undertaken
per lure tested and sprayed trees were always at least
15 m apart from each other. Two parameters were

recorded: 1) to assess the speed of the effect of spot
sprays on B. minax, dead ßies on the sprayed trees
were counted 40 min after the solutions had been

1279

applied, and 2) to evaluate the total number of ßies
killed on the sprayed trees, spot sprays were checked
for dead ßies every day until none were observed.
Overall effectiveness of sprays was assessed according
to the lures used. The number of male and female
killed on spot sprays were analyzed using a generalized linear model (Proc Genmod; SAS Institute 1999)
with Lure and Sex as factors, and interaction between
these two factors was also tested.
Results
Attractiveness of the Lures. The statistical results
are summarized in Table 1A. Overall, a total of 8,894
B. minax adults were ensnared during the season
(4,340 females and 4,554 males). The sex of ßies
trapped was not a driving factor (no signiÞcant Sex
factor). The various lures trapped males and females
in a similar way (no signiÞcant interaction between
Sex and Lure factors) and regardless of date (no signiÞcant interaction between Date and Sex factors).
Lures varied strongly in their attractiveness (i.e., signiÞcant Lure factor, P Ͻ 0.001). Overall, traps with the
H-protein bait trapped much more B. minax ßies than
traps with other lures (Fig. 1). In addition, the SVW
mixture proved to be more efÞcient than Torula yeast
and Jufeng attractant lures. The number of B. minax
ßies also varied according to the date (signiÞcant Date
factor) (Table 1A) with ßies being trapped from midMay through early August and a peak was observed on

8 June (Fig. 1). It reßected the seasonal phenology of
B. minax in the studied area. However, this peak was
more marked in the case of the H-protein bait than for
the other lures, although the SVW mixture showed
also a peak to a lesser extent. For example, the number

Fig. 1. Mean number of B. minax ßies (mean Ϯ SEM) captured per trap weekly using various lures (H-protein bait, SVW,
torula yeast, and Jufeng attractant) in experimental orchards in Hubei province, China.


1280

JOURNAL OF ECONOMIC ENTOMOLOGY

Vol. 105, no. 4

Fig. 2. Female ßies of B. minax in different ovarian development situations ensnared in traps by H-protein bait solution
by date in citrus orchards in the Hubei province, China.

of ßies trapped by H-protein bait traps increased much
more than those trapped by the SVW mixture when ßy
density increased in the orchard, hence signiÞcant
interaction between the Lure and the Date factors
(Table 1A). The torula yeast and Jufeng attractant
lures, that is, the two least efÞcient lures, only trapped
a few ßies throughout the entire season.
B. minax females previously ensnared by H-protein
bait traps were dissected and their sexual maturity was
estimated. Most females trapped by the H-protein bait
were still immature and semimature until mid-June

(Fig. 2). The proportion of fully mature females increased from Ϸ5% in mid-June to almost 90% in late
June. All females trapped from early July until the end

of the test were sexually mature. There was a positive
signiÞcant relationship between the number of female
ßies trapped by the H-protein bait for immature (Fig.
3; R2 ϭ 0.91, F9 ϭ 89.88, P Ͻ 0.001), semimature
(R2 ϭ 0.61, F9 ϭ 13.60, P ϭ 0.005) but not for fully
mature ßies (R2 ϭ 0.01, F9 ϭ 0.12, P ϭ 0.733) (Fig. 3).
Effectiveness of the Lures for Spot Sprays. The
number of females and males killed within 40 min
varied signiÞcantly according to the lures used in
spot sprays (Fig. 4, signiÞcant Lure factor, Table
1B). Overall, during the Þrst 40 min after spray
applications, a similar number of B. minax males and
females were killed on sprayed trees (no signiÞcant
Sex factor) and there was no interaction between

Fig. 3. Relationship between sexual maturity stage (immature, semimature, and mature) ovarian development of females
of B. minax ßies trapped in H-protein bait traps and the numbers of ßies trapped by these traps. Regression lines are included
for the three relationships tested.


August 2012

ZHOU ET AL.: ATTRACTIVE LURES FOR THE FRUIT FLY Bactrocera minax

1281

Fig. 4. Mean number (ϮSEM) of B. minax ßies killed in spot sprays (lure ϩ trichlorphon) within 40 min after application.

Lures tested were H-protein bait, GF-120, SVW mixture, Jufeng attractant, and control (i.e., water).

Lure and Sex factors. Nonetheless, more males than
females tend to be killed in the speciÞc case of
H-protein bait spot sprays, but the results were not
statistically signiÞcant (Fig. 4).
The total number of B. minax females and males
killed on spot sprays (i.e., until dead ßies were no
longer observed on sprayed trees) differed signiÞcantly according to lures used (signiÞcant Lure factor,
Fig. 5, Table 1C). Spot sprays with H-protein bait
ensnared more females than the GF-120, the SVW

mixture, the Jufeng attractant and control spot sprays.
By contrast to what was observed within the Þrst 40
min after initial spot sprays application, signiÞcantly
more males than females were trapped (signiÞcant Sex
factor). The interaction between the two factors was
not signiÞcant, that is, H-protein bait, GF-120 and
SVW mixture spot sprays showed the same trend
(though the less efÞcient Jufeng attractant and the
control sprays did not show this difference in numbers
of male and female trapped).

Fig. 5. Mean total number (ϮSEM) of B. minax ßies killed in spot sprays (lure ϩ trichlorphon) until dead ßies found
dropped to zero (daily). Lures tested were H-protein bait, GF-120, SVW mixture, Jufeng attractant, and control (i.e.,
water).


1282

JOURNAL OF ECONOMIC ENTOMOLOGY
Discussion

Many ßies in the Bactrocera genus are major pests
around the world (Vargas et al. 2009, Daane and Johnson 2010, Pascual et al. 2010, Han et al. 2011, Benelli et
al. 2012, Canale and Benelli 2012) but B. minax is
considered a major pest particularly in Asia. The Sterile Insect Technique (SIT) was used in the 1980s to
manage this pest in the Guizhou province in China;
this method achieved some success (Wang and Luo
1995). However, it relied mainly on government heavy
investment in mass-rearing facilities and the widespread release of sterile insects. In the current study,
we demonstrated that the H-protein bait may be a
promising lure to monitor this ßy in Citrus orchards in
China and moreover to manage this pest when used in
spot sprays. The H-protein bait was the most attractive
lure to both sexes of B. minax; not only in traps but also
in spot sprays. Other lures tested, including the wellknown GF-120, proved to be less efÞcient than the
H-protein bait in attracting B. minax.
Various lures including fruit-derived, food-based,
visual, and pheromone will continue to be an integral
part of the management of tephritid pests. For a given
tephritid pest, the attraction and responses to the lures
are completely different. For example, ßies in the
Bactrocera genus depend much more on olfactory
lures compared with Rhagoletis ßies, which seem to
rely on visual cues (Prokopy and Papaj 2000). The
hydrolyzed proteins were used as baits to attract fruit
ßies, but comparisons of several proteins indicated
that hydrolyzed torula yeast was superior in attracting
Anastrepha spp. (Epsky et al. 1993). In this case, it is

necessary to assess the effect of the protein bait lures
on each target species before commercial use. Unfortunately, very few studies are available about the response of lures in B. minax. Drew et al. (2006) reported
the attractiveness of various combinations of colors
and shapes to B. minax in Bhutan. Our study is the Þrst
to conÞrm that B. minax can have signiÞcant response
to an enzymatical-hydrolyzed protein bait. By quantifying attraction and responses of B. minax compared
with other commercially used lures, a breakthrough
has been made in understanding the H-protein bait
against B. minax. However, many components of the
protein baits (e.g., protein concentrations, pH, and
ammonia concentrations) also likely greatly inßuence
attractiveness for B. minax. Thus, bearing in mind the
aforementioned factors, it would be worth developing
improved lures when furthering work on the Chinese
citrus fruit ßy.
In this Þeld study, we showed that the H-protein
bait was very effective on B. minax both when used in
traps and for spot sprays. However, it is worth mentioning that proteins-based lures used in traps may
perform differently than when sprayed on plant foliage. For example, some proteins sprayed on plants
may be phytotoxic and thus may have to be diluted
before potential use as spot sprays (Mangan et al.
2006), and protein mixtures containing ammonium
acetate may not always work well for trapping purposes (Robacker et al. 1996).

Vol. 105, no. 4

The response of fruit ßies to protein baits is known
to depend greatly on their feeding status and sexual
development (Robacker 1991, Cornelius et al. 2000,
Miller et al. 2004). Our Þeld trials suggested that most

of the B. minax adult ßies ensnared by H-protein bait
traps occurred before late June, that is, about 1 mo
after adult ßies emerged according to the phenology
of B. minax in Hubei province (C.Y.N., unpublished
data). This need for proteins in female ßies may be
exploited as they are likely to be particularly attracted
by protein sources early in the season (Health et al.
1993).
The highest number of female ßies trapped per
week occurred on 8 June and it drastically decreased
on 6 July. Five percent of females trapped on 15 June
were sexually mature whereas almost 96% of females
in traps with H-protein bait solutions in early July
were fully sexually mature (many mature eggs). When
females become sexually mature they need proteins
for the development of eggs and therefore Þnding
protein sources is of primary importance (Mangan
2003, Perez-Staples et al. 2007). Studies reported that
females deprived of nitrogenous food are more attracted by protein bait (Prokopy et al. 1992, 2003;
Vargas et al. 2002; Rousse et al. 2005; Miller et al. 2004).
Our results support the hypothesis that responses of B.
minax females were affected by protein requirement as demonstrated in other Bactrocera species
(Robacker 1991, Miller et al. 2004, Perez-Staples et al.
2007, Mangan 2009). Investigations of ovarian development on B. minax females revealed that ovarian
development appears to be a reliable indicator of sexual maturity, and protein was a vital component of
ovarian development for female B. minax. In laboratory conditions, when B. minax females were fed a full
diet (including proteins provided ad libitum), they
reach sexual maturity and complete the reproduction
phase in Ϸ25 d (C.Y.N., unpublished data). It is also
consistent with the low number of sexually mature

females trapped from late June as ßies no longer
needed much protein (when oogenesis was Þnished).
In our Þeld trials, traps baited with protein lures
captured more nontarget insects than other lures because these insects are attracted to the odors of protein bait (Thomas 2003). The nontarget insects captured were mainly the species belonging to Diptera,
Hymenoptera, and few Coleoptera and Lepidoptera,
such as scarabs and moths. For Diptera, a number of
Muscidae, Calliphoridae, and Sarcophagidae were
trapped. Some beneÞcial insects, like honey bees and
other wild bees, were trapped in small numbers. When
protein bait was used in sprays, these pollinators may
feed on the bait and be affected or even killed by
insecticides (Thomas and Mangan 2005, Desneux et al.
2007). This problem associated with the use of pesticides in bait spray can be solved by using insecticides
which are less toxic on nontarget organisms. Bait
sprays with new types of insecticide that are less harmful to nontarget organisms have been used successfully
in Þeld trials against Anastrepha ludens (Loew), A.
obliqua (Macquart), A. suspensa (Loew), Bactrocera
cucurbitae (Coquillett), Bactrocera dorsalis (Hendel),


August 2012

ZHOU ET AL.: ATTRACTIVE LURES FOR THE FRUIT FLY Bactrocera minax

and Ceratitis capitata (Wiedemann) (Burns et al. 2001,
Moreno et al. 2001, Vargas et al. 2003, Prokopy et al.
2003, Stark et al. 2004, Thomas and Mangan 2005,
Pin˜ ero et al. 2009b). In fact, the new toxicants that are
noticeably different to traditional pesticides are less
likely to kill adults through contact, but instead must

be ingested before producing maximum toxic effect
(DowElanco 1994, Vargas et al. 2002).
The results of the current study may be useful in
optimizing IPM programs for the control of B. minax
in China. Previous IPM programs developed against
Bactrocera ßies hinted at the importance of efÞcient
lures in IPM packages for successful management. For
example, a wide-scale successful IPM program against
B. dorsalis was implemented in Hawaii in 2000 and use
of protein bait sprays and spot sprays (protein baitbased) targeting female ßies was proven to be a keystone component of such IPM (Mau et al. 2007; Vargas
and Prokopy. 2006; Pin˜ ero et al. 2009a,b). Using Hprotein bait has great efÞciency potential when implemented in IPM programs against B. minax, for two
main reasons. Firstly, H-protein bait showed great
efÞciency in mass trapping as well as in spot spray
targeting B. minax adults. In practice, adults are caught
or killed before they became sexually mature, that is,
before the ßies effectively start to lay eggs on or in
fruits and thus it could prevent any adverse effects on
fruits by larvae. Secondly, the speciÞc phenology of B.
minax, that is, univoltine species, may allow IPM programs to focus most of management efforts for a relatively short time span in the year; from the emergence of ßies and until they become sexually mature.
The main sanitation measures in Citrus orchards, for
example, collection and removal of all fallen infested
fruits at the end of the fruit season combined with
H-protein bait mass trapping and spot sprays can
largely suppress the B. minax populations and therefore ensure a successful IPM program. In addition, the
implementation of such a strategy may be cost-effective and environmentally sound, it would particularly
help to reduce pesticide applications and possible negative effects on beneÞcial arthropods (Desneux et al.
2006, 2007; Han et al. 2010) and on associated ecosystem services (Lu et al. 2012).
The data presented here strongly showed that Hprotein bait can be an effective lure for both female
and male B. minax in citrus orchards during the fruiting season. Protein bait attracted more B. minax adults
than other attractants in this study. Further studies are

required to enhance the formulation and the spray
concentration; moreover, this protein bait can be used
in combination with attract-and-kill devices.

Acknowledgments
The authors thank the International Atomic Energy
Agency Research (Grant no. 16015) for funding to C.Y.N. and
the Plant Health & Environment Department of INRA for
funding to N.D.

1283

References Cited
Benelli, G., G. Bonsignori, C. Stefanini, and A. Canale. 2012.
Courtship and mating behaviour in the fruit ßy parasitoid
Psyttalia concolor (Sze´ pligeti) (Hymenoptera: Braconidae): the role of wing fanning. J. Pest Sci. 85: 55Ð 63.
Burns, R. E., D. L. Harris, D. S. Moreno, and J. E. Eger. 2001.
EfÞcacy of spinosad bait sprays to control Mediterranean
and Caribbean fruit ßies in commercial citrus in Florida.
Fla. Entomol. 84: 672Ð 678.
Canale, A., and G. Benelli. 2012. Impact of mass-rearing on
the host seeking behaviour and parasitism by the fruit ßy
parasitoid Psyttalia concolor (Sze´ pligeti) (Hymenoptera:
Braconidae). J. Pest Sci. 85: 65Ð74.
Christenson, L. D., and R. H. Foote. 1960. Biology of fruit
ßies. Annu. Rev. Entomol. 5: 171Ð192.
Cornelius, L. M., L. Nergel, J. J. Duan, and R. H. Messing.
2000. Responses of female oriental fruit ßies (Diptera:
Tephritidae) to protein and host fruit odor in Þeld cage
and open Þeld tests. Environ. Entomol. 29: 14 Ð19.

Daane, K. M., and M. W. Johnson. 2010. Olive fruit ßy:
managing an ancient pest in modern times. Annu. Rev.
Entomol. 55: 151Ð169.
Desneux, N., A. Decourtye, and J. M. Delpuech. 2007. The
sublethal effects on beneÞcial arthropods. Annu. Rev.
Entomol. 52: 81Ð106.
Desneux, N., R. Ramirez-Romero, and L. Kaiser. 2006. Multi
step bioassay to predict recolonization potential of
emerging parasitoids after a pesticide treatment. Environ.
Toxicol. Chem. 25: 2675Ð2682.
Dorji, C., A. R. Clarke, R.A.I. Drew, B. S. Fletcher, P. Loday,
K. Mahat, S. Raghu, and M. C. Romig. 2006. Seasonal
phenology of Bactrocera minax (Diptera: Tephritidae) in
western Bhutan. B. Entomol. Res. 96: 531Ð538.
DowElanco. 1994. Spinosad technical guide. DowElanco,
Indianapolis, IN.
Drew, R.A.I. 1979. The genus Dacus Fabricius (Diptera: Tephritidae): two new species from northern Australia and
a discussion of some subgenera. J. Aust. Entomol. Soc. 18:
71Ð 80.
Drew, R.A.I., A. C. Courtice, and D. S. Teakle. 1983. Bacteria as a natural source of food for adult fruit ßies (Diptera: Tephritidae). Oecologia 60: 279 Ð284.
Drew, R.A.I., C. Dorji, M. C. Romig, and P. Loday. 2006.
Attractiveness of various combinations of colors and
shapes to females and males of Bactrocera minax (Diptera:
Tephritidae) in a commercial mandarin grove in Bhutan.
J. Econ. Entomol. 99: 1651Ð1656.
Epsky, N. D., R. R. Heath, J. M. Sivinski, C. O. Calkins, R. M.
Baranowski, and A. H. Fritz. 1993. Evaluation of protein
bait formulations for the Caribbean fruit ßy (Diptera:
Tephritidae). Fla. Entomol. 76: 626 Ð 635.
Han, P., C. Y. Niu, C. L. Lei, J. J. Cui, and N. Desneux. 2010.

Use of an innovative T-tube maze assay and the proboscis
extension response assay to assess sublethal effects of GM
products and pesticides on learning capacity of the honey
bee Apis mellifera L. Ecotoxicology 19: 1612Ð1619.
Han, P., X. Wang, C. Y. Niu, Y. C. Dong, and N. Desneux.
2011. The population dynamics, phenology and overwintering of Bactrocera dorsalis (Diptera: Tephritidae) in
Hubei province, China. J. Pest Sci. 84: 289 Ð395.
Heath, R. R., N. D. Epsky, P. J. Landolt, and J. Sivinski. 1993.
Development of attractants for monitoring Caribbean
fruit ßies (Diptera: Tephritidae). Fla. Entomol. 76: 233Ð
244.
Hendrichs, J., G. Franz, and P. Rendon. 1995. Increased
effectiveness and applicability of the sterile insect technique through male-only releases for control of Mediter-


1284

JOURNAL OF ECONOMIC ENTOMOLOGY

ranean fruit ßies during fruiting seasons. J. Appl. Entomol.
119: 371Ð377.
Lu, Y. H., K. M. Wu, Y. Y. Jiang, Y. Y. Guo, and N. Desneux.
2012. Widespread adoption of Bt cotton and insecticide
decrease promotes biocontrol services. Nature. (in
press). (doi:10.1038/nature11153).
Mangan, R. L. 2003. Adult diet and male-female contact
effects on female reproductive potential in Mexican fruit
ßy (Anastrepha ludens Loew) (Diptera: Tephritidae). J.
Econ. Entomol. 96: 341Ð347.
Mangan, R. L. 2009. Effects of bait age and prior protein

feeding on cumulative tie-dependent mortality of Anastrepha ludens (Diptera: Tephritidae) exposed to GF-120
spinosad baits. J. Econ. Entomol. 102: 1157Ð1163.
Mangan, R. L., D. S. Moreno, and G. D. Thompson. 2006.
Bait dilution, spinosad concentration, and efÞcacy of GF120 based fruit ßy sprays. Crop. Prot. 25: 125Ð133.
Mau, R.F.L., E. B. Jang, and R. I. Vargas. 2007. The Hawaii
fruit ßy area-wide fruit ßy pest management programme:
inßuence of partnership and a good education programme, pp. 671Ð 683. In M.J.B. Vreysen, A. S. Robinson,
and J. Hendrichs (eds.), Area-Wide Control of Insect
Pests: From Research to Field Implementation. Springer,
Dordrecht, The Netherlands.
McQuate, G. T. 2009. Effectiveness of GF-120NF fruit ßy
bait as a suppression tool for Bactrocera latifrons (Diptera:
Tephritidae). J. Appl. Entomol. 133: 444 Ð 448.
Miller, N. W., R. I. Vargas, R. J. Prokopy, and B. E. Mackey.
2004. State-dependent attractiveness of protein bait and
host fruit odor to Bactrocera cucurbitae (Diptera: Tephritidae) females. Ann. Entomol. Soc. Am. 97: 1063Ð
1068.
Moreno, D. S., H. Celedonio, R. L. Mangan, J. L. Zavala, and
P. Montoya. 2001. Field evaluation of a phototoxic dye,
phloxine B, against three species of fruit ßies (Diptera:
Tephritidae). J. Econ. Entomol. 94: 1419 Ð1427.
Newell, W. 1936. Progress report on the Key West (Florida)
fruit ßy eradication project. J. Econ. Entomol. 29: 116 Ð
120.
Pascual, S., G. Cobos, E. Seris, and M. Gonzalez-Nunez. 2010.
Effects of processed kaolin on pests and non-target arthropods in a Spanish olive grove. J. Pest Sci. 83: 121Ð133.
Perez-Staples, D., V. Prabhu, and P. W. Taylor. 2007. Postteneral protein feeding enhances sexual performance of
Queensland fruit ßies. Physiol. Entomol. 32: 225Ð232.
Pin˜ ero, J. C., R.F.L. Mau, G. T. McQuate, and R. I. Vargas.
2009a. Novel bait stations for attract-and-kill of pestiferous fruit ßies. Entomol. Exp. Appl. 133: 208 Ð216.

Pin˜ ero, J. C., R.F.L. Mau, and R. I. Vargas. 2009b. Managing
oriental fruit ßy (Diptera: Tephritidae), with spinosadbased protein bait sprays and sanitation in papaya orchards in Hawaii. J. Econ. Entomol. 102: 1123Ð1132.
Prokopy, R. J., N. W. Miller, J. C. Pin˜ ero, J. D. Barry, L. C.
Tran, L. Oride, and R. I. Vargas. 2003. Effectiveness of
GF-120 fruit ßy bait spray applied to border area plants
for control of melon ßies (Diptera: Tephritidae). J. Econ.
Entomol. 96: 1485Ð1493.
Prokopy, R. J., D. R. Papaj, J. Hendrichs, and T.T.Y. Wong.
1992. Behavioral responses of Ceratitis capitata ßies to
bait spray droplets and natural food. Entomol. Exp. Appl.
64: 247Ð257.
Prokopy, R. J., and D. R. Papaj. 2000. Behavior of ßies of the
genera Rhagoletis, Zonosemata, and Carpomya (Trypetinae: Carpomyina), pp. 219 Ð252. In M. Aluja and A. L.

Vol. 105, no. 4

Norrbom (eds.), Fruit Flies (Tephritidae): Phylogeny
and Evolution of Behavior. CRC, Boca Raton, FL.
Robacker, D. C. 1991. SpeciÞc hunger in Anastrepha ludens
(Diptera: Tephritidae): effects on attractiveness of proteinaceous and fruit-derived lures. Environ. Entomol. 20:
1680 Ð1686.
Robacker, D. C., D. S. Moreno, and A. B. Demilo. 1996.
Attractiveness to Mexican fruit ßies of combinations of
acetic acid with ammonium/amino attractants with emphasis on effects of hunger. J. Chem. Ecol. 22: 499 Ð511.
Rousse, P., P. F. Duyck, S. Quilici, and P. Ryckewaert. 2005.
Adjustment of Þeld cage methodology for testing food
attractants for fruit ßies. Ann. Entomol. Soc. Am. 98:
402Ð 408.
SAS Institute. 1999. SAS/Stat userÕs guide, release 8. ed. SAS
Institute, Cary, NC.

Stark, J. D., R. I. Vargas, and N. W. Miller. 2004. Toxicity of
spinosad in protein bait to three economically important
tephritid fruit ßy species (Diptera: Tephritidae) and their
parasitoids (Hymenoptera: Braconidae). J. Econ. Entomol. 97: 911Ð915.
Steiner, L. F. 1952. Fruit ßy control in Hawaii with poisonbait sprays containing protein hydrolysates. J. Econ. Entomol. 45: 838 Ð 843.
Stonehouse, J., M. Afzal, Q. Zia, J. Mumford, A. Poswal, and
R. Mahmood. 2002. “Single-killing-point” Þeld assessment of bait and lure control of fruit ßies (Diptera: Tephritidae) in Pakistan. Crop. Prot. 21: 651Ð 659.
Thomas, D. B., and R. L. Mangan. 2005. Nontarget impact of
spinosad GF-120 bait sprays for control of the Mexican
fruit ßy (Diptera: Tephritidae) in Texas citrus. J. Econ.
Entomol. 98: 1950 Ð1956.
Thomas, D. B. 2003. Nontarget insects captured in fruit ßy
(Diptera: Tephritidae) surveillance traps. J. Econ. Entomol. 96: 1732Ð1737.
Vargas, R. I., and R. J. Prokopy. 2006. Attraction and feeding
responses of melon ßies and oriental fruit ßies (Diptera:
Tephritidae) to various protein baits with and without
toxicants. Proc. Hawaiian Entomol. Soc. 38: 49 Ð 60.
Vargas, R. I., N. W. Miller, and R. J. Prokopy. 2002. Attraction and feeding responses of Mediterranean fruit ßy and
a natural enemy to protein baits laced with two novel
toxins, phloxine B and spinosad. Entomol. Exp. Appl. 102:
273Ð282.
Vargas, R. I., N. W. Miller, and J. D. Stark. 2003. Field trials
of spinosad as a replacement for naled, DDVP, and malathion in methyl eugenol and cue-lure bucket traps to
attract and kill male oriental fruit ßies and melon ßies
(Diptera: Tephritidae) in Hawaii. J. Econ. Entomol. 96:
1780 Ð1785.
Vargas, R. I., J. C. Pin˜ ero, R.F.L. Mau, J. D. Stark, and M.
Hertlein. 2009. Attraction and mortality of oriental fruit
ßies (Diptera: Tephritidae) to SPLAT-MAT-methyl eugenol with spinosad. Entomol. Exp. Appl. 131: 286 Ð293.
Wang, X. J., and L. Y. Luo. 1995. Research progress in the

Chinese citrus fruit ßy. Entomol. Knowl. 32: 310 Ð315.
Yang, P., J. R. Carey, and R. V. Dowell. 1994. Tephritid fruit
ßies in China: historical background and current status.
Pan-Pac. Entomol. 70: 159 Ð167.
Yee, W. L., and P. S. Chapman. 2005. Effects of GF-120
concentrations on attraction, feeding, mortality, and control of Rhagoletis indifferens (Diptera: Tephritidae). J.
Econ. Entomol. 98: 1654 Ð1663.
Received 15 January 2012; accepted 29 May 2012.