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

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.

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 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

https://doi.org/10.1603/EC12020

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.

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,1CHANG-YING NIU,1,2PENG HAN,1,3

ANDNICOLAS DESNEUX3

J Econ Entomol 105(4): 1277Ð1284 (2012); DOI: http://dx.doi.org/10.1603/EC12020

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 signiÞ-cantly 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

(Ender-lein) (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

consider-able economic damage by making the fruits improper

for merchandizing (Yang et al 1994) Therefore,

ef-fective Integrated Pest Management (IPM) programs

are required for areas where the citrus production is

a major economic issue

Traps, attractive lures, and mass trapping tech-niques 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 ap-proach combining olfactory attractants and baits sprayed with insecticides, that is, lures⫹ 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 erad-icating 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,

1 Hubei Insect Resources Utilization and Sustainable Pest

Manage-ment Key Laboratory, College of Plant Science & Technology,

Hua-zhong Agricultural University, Wuhan 430070, China.

2 Corresponding author, e-mail: niuchy2004@yahoo.com.cn.

3 French National Institute for Agricultural Research (INRA), 400

route des chappes, 06903 Sophia-Antipolis, France.

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

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,

farm-ers frequently rely on spraying sugar-vinegar-wine

mixture with insecticides to manage the pest (Wang

and Luo 1995) Though most fruit ßies can be

moni-tored by traps baited with sexual pheromone and

para-pheromones, 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

avail-able as lure for B minax in China though its efÞcacy

has never been clearly demonstrated In our

labora-tory, a new protein bait has recently been developed

to control B minax It is based on an enzymatically

hydrolyzed protein produced by the industrial

pro-cessing of beer yeast This hydrolyzed protein bait

(hereafter named H-protein bait) also includes

feed-ing stimulants, orange juice and brown sugar, as well

as a chemical attractant ammonium acetate

In 2010, in Þeld conditions we assessed the

effec-tiveness of the H-protein bait and several other

com-mercially 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-vin-egar-wine (SVW) mixture, and Jufeng as attractant for

B minax, and 2) we studied how various lures

(H-protein 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

insecti-cide 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 ⬇1,000 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

rel-ative humidity during the experiments were 27.2⫾

0.3⬚C, 67.5 ⫾ 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: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 periodi-cally, 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⫻ 3 cm window 8

cm from the bottom of the bottle) This design en-snares 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 at-tached 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⫻ 9 ⫽ 36 traps were used every week Once a week, when traps were checked and ßies were removed, the traps were there-after 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 sub-sequently dissected to estimate their sexual maturity under a stereomicroscope (Nikon Inc., SMC-10, Ja-pan) and placed into three categories: immature (ova-ries small and no eggs present), semimature (ova(ova-ries 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

mi-nax 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 lure-insecticide solutions, that is, spot sprays, were pre-pared: 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

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

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 general-ized 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 sig-niÞcant interaction between Date and Sex factors) Lures varied strongly in their attractiveness (i.e.,

sig-niÞcant Lure factor, P⬍ 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 mid-May 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

Table 1 Statistics from the generalized linear model used to

analyze the numbers of flies

Source of variationa df ␹ 2 P value

A: Lure attractiveness

Lure ⫻ Date 30 654.01 ⬍0.001

Date ⫻ Sex 10 16.34 0.090

B: Spot spraysÑimmediate effect

C: Spot spraysÑtotal effect

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.

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

in-creased from⬇5% 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 ⫽ 0.91, F9 ⫽ 89.88, P ⬍ 0.001), semimature (R2⫽ 0.61, F9⫽ 13.60, P ⫽ 0.005) but not for fully mature ßies (R2⫽ 0.01, F9⫽ 0.12, P ⫽ 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 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

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

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

signiÞ-cantly 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 4 Mean number (⫾SEM) of B minax ßies killed in spot sprays (lure ⫹ trichlorphon) within 40 min after application.

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

Fig 5 Mean total number (⫾SEM) of B minax ßies killed in spot sprays (lure ⫹ 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)

Many ßies in the Bactrocera genus are major pests

around the world (Vargas et al 2009, Daane and

John-son 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

Ster-ile 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

wide-spread 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

well-known 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

Unfor-tunately, very few studies are available about the

re-sponse 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

quan-tifying 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

men-tioning that proteins-based lures used in traps may

perform differently than when sprayed on plant

foli-age 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

pur-poses (Robacker et al 1996)

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 at-tracted 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

require-ment as demonstrated in other Bactrocera species

(Robacker 1991, Miller et al 2004, Perez-Staples et al

2007, Mangan 2009) Investigations of ovarian

devel-opment on B minax females revealed that ovarian

development appears to be a reliable indicator of sex-ual maturity, and protein was a vital component of

ovarian development for female B minax In labora-tory 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⬇25 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 be-cause these insects are attracted to the odors of pro-tein bait (Thomas 2003) The nontarget insects cap-tured 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 pesti-cides in bait spray can be solved by using insectipesti-cides which are less toxic on nontarget organisms Bait sprays with new types of insecticide that are less harm-ful to nontarget organisms have been used successharm-fully

in Þeld trials against Anastrepha ludens (Loew), A.

obliqua (Macquart), A suspensa (Loew), Bactrocera cucurbitae (Coquillett), Bactrocera dorsalis (Hendel),

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

bait-based) targeting female ßies was proven to be a

key-stone component of such IPM (Mau et al 2007; Vargas

and Prokopy 2006; Pin˜ero et al 2009a,b) Using

H-protein bait has great efÞciency potential when

im-plemented 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

pro-grams to focus most of management efforts for a

rel-atively short time span in the year; from the

emer-gence 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

there-fore ensure a successful IPM program In addition, the

implementation of such a strategy may be

cost-effec-tive and environmentally sound, it would particularly

help to reduce pesticide applications and possible

neg-ative effects on beneÞcial arthropods (Desneux et al

2006, 2007; Han et al 2010) and on associated

ecosys-tem services (Lu et al 2012)

The data presented here strongly showed that

H-protein bait can be an effective lure for both female

and male B minax in citrus orchards during the

fruit-ing 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

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:

Braconi-dae): the role of wing fanning J Pest Sci 85: 55Ð63

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: 672Ð678

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: 14Ð19

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:

Te-phritidae): two new species from northern Australia and

a discussion of some subgenera J Aust Entomol Soc 18: 71Ð80

Drew, R.A.I., A C Courtice, and D S Teakle 1983

Bac-teria as a natural source of food for adult fruit ßies (Dip-tera: Tephritidae) Oecologia 60: 279Ð284

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: 626Ð635

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

overwin-tering of Bactrocera dorsalis (Diptera: Tephritidae) in

Hubei province, China J Pest Sci 84: 289Ð395

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 tech-nique through male-only releases for control of

Mediter-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

Anas-trepha 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

GF-120 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

pro-gramme, pp 671Ð683 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: 444Ð448

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:

Te-phritidae) 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: 1419Ð1427

Newell, W 1936 Progress report on the Key West (Florida)

fruit ßy eradication project J Econ Entomol 29: 116Ð

120

Pascual, S., G Cobos, E Seris, and M Gonzalez-Nunez 2010.

Effects of processed kaolin on pests and non-target

ar-thropods in a Spanish olive grove J Pest Sci 83: 121Ð133

Perez-Staples, D., V Prabhu, and P W Taylor 2007

Post-teneral 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

pestifer-ous fruit ßies Entomol Exp Appl 133: 208Ð216

Pin˜ero, J C., R.F.L Mau, and R I Vargas 2009b Managing

oriental fruit ßy (Diptera: Tephritidae), with

spinosad-based protein bait sprays and sanitation in papaya

or-chards 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

(Trypeti-nae: Carpomyina), pp 219Ð252 In M Aluja and A L.

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 pro-teinaceous and fruit-derived lures Environ Entomol 20: 1680Ð1686

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 em-phasis on effects of hunger J Chem Ecol 22: 499Ð511

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: 402Ð408

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 Ento-mol 97: 911Ð915

Steiner, L F 1952 Fruit ßy control in Hawaii with

poison-bait sprays containing protein hydrolysates J Econ En-tomol 45: 838Ð843

Stonehouse, J., M Afzal, Q Zia, J Mumford, A Poswal, and

R Mahmood 2002 “Single-killing-point” Þeld

assess-ment of bait and lure control of fruit ßies (Diptera: Te-phritidae) in Pakistan Crop Prot 21: 651Ð659

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: 1950Ð1956

Thomas, D B 2003 Nontarget insects captured in fruit ßy

(Diptera: Tephritidae) surveillance traps J Econ Ento-mol 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: 49Ð60

Vargas, R I., N W Miller, and R J Prokopy 2002

Attrac-tion 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 mal-athion 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: 1780Ð1785

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 eu-genol with spinosad Entomol Exp Appl 131: 286Ð293

Wang, X J., and L Y Luo 1995 Research progress in the

Chinese citrus fruit ßy Entomol Knowl 32: 310Ð315

Yang, P., J R Carey, and R V Dowell 1994 Tephritid fruit

ßies in China: historical background and current status Pan-Pac Entomol 70: 159Ð167

Yee, W L., and P S Chapman 2005 Effects of GF-120

concentrations on attraction, feeding, mortality, and

con-trol of Rhagoletis indifferens (Diptera: Tephritidae) J.

Econ Entomol 98: 1654Ð1663

Received 15 January 2012; accepted 29 May 2012.