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Host discrimination and superparasitism in wild and mass-rearedDiachasmimorpha longicaudata (Hym.: Braconidae) femalesP. I. González a; P. Montoya b; G. Pérez-Lachaud a; J. Cancino b; P. Liedo a

a El Colegio de la Frontera Sur, Tapachula, Chiapas, Mexico b Programa Moscafrut, SAGARPA-IICA,Tapachula, Chiapas, Mexico

Online publication date: 14 December 2009

To cite this Article González, P. I., Montoya, P., Pérez-Lachaud, G., Cancino, J. and Liedo, P.(2010) 'Host discriminationand superparasitism in wild and mass-reared Diachasmimorpha longicaudata (Hym.: Braconidae) females', BiocontrolScience and Technology, 20: 2, 137 — 148To link to this Article: DOI: 10.1080/09583150903437266URL: http://dx.doi.org/10.1080/09583150903437266

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RESEARCH ARTICLE

Host discrimination and superparasitism in wild and mass-rearedDiachasmimorpha longicaudata (Hym.: Braconidae) females

P.I. Gonzaleza, P. Montoyab*, G. Perez-Lachauda, J. Cancinob and P. Liedoa

aEl Colegio de la Frontera Sur, Carretera Antiguo Aeropuerto Km 2.5, Tapachula, Chiapas30700, Mexico; bPrograma Moscafrut, SAGARPA-IICA, Av. Central Poniente 14, Col.

Centro, Tapachula Chiapas 30700, Mexico

(Received 11 May 2009; returned 22 July; accepted 23 October 2009)

We compared the oviposition behavior and host discrimination ability of wild andmass-reared Diachasmimorpha longicaudata (Hymenoptera: Braconidae) femalesparasitizing Anastrepha ludens (Diptera: Tephritidae) larvae. Both kinds ofparasitoid females were presented simultaneously with parasitized and non-parasitized larvae in choice tests, and their superparasitism performance wasevaluated under a mass-rearing situation. At the time of the test, D. longicaudatahad 156 generations under mass-rearing conditions. Our goal was to determinethe effect of the mass-rearing process on the foraging decisions of this species.One of the primary findings was the apparent ubiquity of superparasitism byD. longicaudata females. Both types of females showed similar patterns in each ofthe phases of oviposition behavior evaluated. The only notable differences wereamong the percentages of transition between behaviors, mainly related to theintensity with which each activity was performed. Under a mass-rearing situation,both strains of females had a similar tendency to increase superparasitism (i.e.,number of oviposition scars per puparium and the proportion of superparasitizedlarvae) over time. The mass-rearing process appears to have induced the selectionof more aggressive, fertile and precocious females. Despite these observations, weconcluded that the process of adaptation to mass-rearing conditions has notsubstantially influenced the foraging and ovipositional behaviors in this species.

Keywords: Diachasmimorpha longicaudata; superparasitism; foraging decisions;mass rearing

Introduction

Superparasitism � a female laying eggs in an already parasitized host � is a

phenomenon frequently observed in the laboratory (e.g., van Alphen and Visser

1990; Montoya et al. 2003; White and Andow 2008) as well as in nature (van

Lenteren 1981; van Dijken and Waage 1987; Baker, Peulet, and Visser 1990).

Parasitoids used as biocontrol agents are expected to be highly efficient in finding

hosts and to be able to discriminate between parasitized and non-parasitized hosts

(van Lenteren, Bakker, and van Alphen 1978), avoiding superparasitism and

minimizing the time and energy associated with searching behavior (Godfray 1994;

Mackauer 1990).

*Corresponding author. Email: pmontoya@prodigy.net.mx

ISSN 0958-3157 print/ISSN 1360-0478 online

# 2010 Taylor & Francis

DOI: 10.1080/09583150903437266

http://www.informaworld.com

Biocontrol Science and Technology,

Vol. 20, No. 2, 2010, 137�148

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In the past, superparasitism has been attributed to the inability of females to

discriminate between parasitized and non-parasitized hosts, and has been interpreted

as an error by the ovipositing female. However, different authors have stated that

under specific conditions, superparasitism may be an adaptive strategy (see van

Alphen and Visser 1990; Rosenheim and Hongkham 1996; White and Andow 2008),

resulting from a balance between the benefits and the costs of laying an egg in an

already parasitized host. The advantages of superparasitism are an increasedpossibility of producing offspring from a host and the stabilization of host�parasitoid interactions in solitary and gregarious parasitoids (van Alphen 1988;

van Alphen and Visser 1990).

According to Wanjberg, Pizzol, and Babault (1989), a propensity for super-

parasitism in Trichogramma maidis Pintureau and Voegele (Hymenoptera: Tricho-

grammatidae) seems to be genetically determined in mass-reared individuals. In

contrast, results by White and Andow (2008) in Macrocentrus grandii (Hymenop-

tera: Braconidae), suggest that host availability and female environmental

perception of conspecific competitors (rather than wasp propensity), determine

self-superparasitism. Other studies have also shown that parasitoids adaptively

superparasitize when perceiving a limitation of host resources (van Alphen and

Visser 1990; Weisser and Houston 1993). Superparasitism studies should be

considered in parasitoid mass-rearing protocols and augmentative field release

biocontrol programs, to ensure production efficiency, to prevent waste of parasitoid

reproductive potential and to infer the performance of the parasitoids in the field.Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae) is a fruit

fly parasitoid considered worldwide as an effective biological control agent for

release purposes (Montoya et al. 2000b, 2007; Ovruski, Colin, Soria, Orono, and

Schliserman 2003). Adult females parasitize a number of second- and third-instar

fruit fly species’ larvae in a wide variety of host fruits (Sivinski, Pinero, and Aluja

2000). The species was originally discovered in the Indo-Philippine region where it

attacked Bactrocera spp. (Wharton and Marsh 1978). In addition to wide-spread

introductions, D. longicaudata has been mass-reared and released by augmentation,

either alone or in combination with sterile male flies for the control of Ceratitis

capitata (Wong et al. 1991) and various Anastrepha spp. (Sivinski et al. 1996;

Montoya et al. 2000b). This species develops as a solitary koinobiont endoparasitoid

of larvae/prepupae and emerges from the host puparium. Unlike some other

tephritid-attacking opiines, D. longicaudata forages extensively for larval hosts in

fallen fruit (Purcell, Jackson, Long, and Batchelor 1994). Female lifespan offspring

production averages 213.494.3 (Gonzalez, Montoya, Perez-Lachaud, Cancino, and

Liedo 2007).While the capacity for host discrimination in this species has been reported

(Lawrence 1988; Montoya et al. 2003), Montoya et al. (2000a) found a strong

tendency in this species toward self-superparasitism in laboratory experiments, even

in the presence of abundant non-parasitized hosts. Recently, Gonzalez et al. (2007)

determined a high incidence of superparasitism in D. longicaudata under mass-

rearing conditions, although superparasitism was correlated with an efficacious

female-biased sex-ratio, and had no negative consequences in any of the life-history

traits studied (longevity, fecundity and flight ability). These results suggest that in

this species superparasitism could be adaptive. However, the incidence of super-

parasitism by wild females in the field is not known.

138 P.I. Gonzalez et al.

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In this work, we compared the oviposition behavior and host discrimination

ability of wild and mass-reared D. longicaudata females attacking Anastrepha ludens

(Loew) (Diptera: Tephritidae) larvae under laboratory conditions. Both kinds

of parasitoid females were presented simultaneously with parasitized and non-

parasitized larvae in choice tests. We video recorded the oviposition behavioral

sequences of both types of females and recorded their preference with respect to the

kind of larvae attacked. We also studied, under a mass-rearing situation, the

superparasitism that both kinds of females exhibited with regard to the host larvae

during five consecutive days. Our goal was to find the effect (if any) that the mass-

rearing process had on the ovipositional foraging behavior of this species.

Materials and methods

Work site and biological material

This work was carried out at the Biological Control Laboratory of the MOSCAFRUT

Program, SAGARPA-IICA in Metapa, Chiapas, Mexico. Puparia of D. longicaudata

and larvae of A. ludens were provided by the MOSCAFRUT mass-rearing facility,

where insects are produced according to the methods described by Cancino (1997) and

Domınguez, Hernandez, and Moreno (1997), respectively. Wild individuals were

obtained from infested mango fruits, Mangifera indica (Anacardiaceae) var. ‘Creole’

collected in the county of Mazapa de Madero, where D. longicaudata was released 10

years ago and was not released thereafter (J. Cancino, personal observation). The

mango fruits were dissected and the larvae were placed in plastic containers with

humid vermiculite to induce pupation. Before emergence, puparia were placed in

mesh-covered wood frame (30�30�30 cm) cages. Adults of both sexes were

maintained in these cages until used for experiments and provided with food

(crystallized honey) to ensure mating. Laboratory conditions were 24928C, 70�80%

RH, and photoperiod of 12 h L:12 h D until adult emergence.

Host discrimination

Host discrimination of wild and mass-reared D. longicaudata females was examined

in choice tests. Experiments basically follow the procedures described by Montoya

et al. (2003) and the criteria proposed by van Alphen and Jervis (1996). Twenty-five

laboratory females and 20 wild females (5�8-days-old and without oviposition

experience) were individually exposed to the simultaneous presence of parasitized

and non-parasitized third instar A. ludens larvae. The parasitized larvae were

obtained by exposing A. ludens larvae to D. longicaudata females 1 day before the

experiments, for 20 min. Using a stereomicroscope we verified the presence of one to

three oviposition scars in each fruit fly larva, which, according to Montoya et al.

(2003) should result in at least one parasitoid egg.

The experimental arena consisted of a 24-cell clear plastic chamber (8.5�12.5 cm,

wide�long). Each individual cell was 1.5�0.5 cm, diameter�deep, where para-

sitized and unparasitized larvae were individually placed in an alternating fashion,

then provided with an abundant semi-synthetic diet. The mesh covering the chamber

was previously impregnated with mango juice, in order to provide some host plant

stimulus to females (Greany, Tumlinson, Chambers, and Boush 1977; Carrasco,

Biocontrol Science and Technology 139

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Montoya, Cruz-Lopez, and Rojas 2005). The behavior of each female was observed

for 30 min. Data on the first larvae chosen, the kind of larvae subsequently attacked,

the total number of ovipositions and oviposition attempts, as well as the duration of

these events, were recorded. All observations were carried out under lab conditions, at24928C temperature and 7095% RH. The behavior of 17 females (eight mass-reared

and nine wild females) was also recorded using a digital video-camera Sony† type

DCR-TRV840, in order to describe and quantify the observed behavior.

The behaviors performed by each female were transcribed sequentially in a first-

order matrix, and the transition frequencies between pairs of behaviors were calcu-

lated to determine their level of importance. Based on the transition frequencies, we

built a flow diagram for each strain of females.

Superparasitism by wild and lab females under a mass-rearing situation

Parasitoids were placed in wood frame, (30�30�30 cm) mesh-covered cages at a 2:1

host larvae:female parasitoid ratio to simulate mass-rearing conditions. Theoviposition units were lids of Petri dishes (10�1 cm, diameter�deep) into which

60 irradiated and unparasitized third instar A. ludens larvae were placed with larval

diet, and then covered with a fine mesh. Females were 5�7-days-old and without

oviposition experience at the beginning of the experiment.

Simultaneously but in independent cages, the host larvae were exposed to the

attack of females from each strain over the course of five consecutive days for 2 h per

day. Oviposition units were renewed every day. The level of superparasitism and its

evolution during the 5 days of larval exposure were determined for each type of female,by counting the number of oviposition scars per puparium per day. As shown by

Montoya et al. (2000a) the number of oviposition scars on the attacked puparia is

strongly correlated with the number of first instar parasitoid larvae found by dissection

(R2� 0.901, see their Figure 3). Seven replicates of this experiment were done.

Statistical analysis

The type of host (parasitized or non-parasitized) chosen by wild and lab reared

females for their first oviposition, and the number of self-superparasitisms were

compared using a Chi-square test. The proportions of wild and lab reared females that

selected non-parasitized larvae for the first five oviposition events in the host

discrimination experiment were compared by applying Fisher’s Exact Test (Zar 1984).Because females attacked a different number of hosts during the 30 min observation

period (see Results), we compared the number of probing events and the number and

duration of ovipositions that females of both types performed on already parasitized

hosts through covariance analyses, with the total number of attempts, the total

number of ovipositions, and the total time each female spent in oviposition as a

covariable for the corresponding analysis. Accurate data on oviposition duration were

available only for the video-recorded females. Superparasitism under the mass-rearing

situation was analyzed by a factorial analysis of variance (strain of female and day ofoviposition as factors); means were separated by the Tukey�Kramer HSD (a�0.05)

procedure. We also performed a simple lineal regression analysis between the number

of scars per puparium and the proportion of superparasitized larvae in successive

days. All statistical analyses were done using Statistica ver. 7 (Statsoft 2004).

140 P.I. Gonzalez et al.

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Results

Host discrimination

While wild females, in their first oviposition attempt selected non-parasitized larvae

more frequently than mass-reared females, this difference was not significant (Table 1;

x2c1, 0.95�2.57, P�0.05). After their first oviposition, wild and lab females attacked

similar numbers of non-parasitized larvae. For example, 55% of mass-reared females

and 60% of wild females selected a non-parasitized larva for their second and third

ovipositions. Moreover, when the first five choices of each female were compared,

there was no statistical difference in the proportions of females of each strain that

selected non-parasitized larvae (Fisher test, P�0.93, 0.74, 0.63, 0.28 and 0.32 for the

first to the fifth ovipositions, respectively).

Females from the mass-rearing facility probed as many hosts as wild females,

irrespective of the type of host attacked (lab females 15.791.3, wild females 15.391.4,

mean9SEM; ANCOVA F1,25�0.04 P�0.84), although the first ones successfully

parasitized a slightly higher number of larvae during the 30 min of observation

(ANOVA, F1,43�6.34, P�0.02; lab females 890.46 vs. wild females 6.4590.38).

There were no statistical differences among the number of ovipositions that females of

the two strains made into already parasitized hosts (ANCOVA, F1, 42�0.06, P�0.81),

or in the mean duration of ovipositions (ANCOVA, F1,14�0.07, P�0.798) (Table 1).It is interesting to note that females of both strains self-superparasitized their

hosts, 28% of mass-reared females (7/25) and 30% of wild females (6/20) self

superparasitized at least one host, with no significant difference between female

types (x2c1, 0.95�0.2, P�0.05). There was no significant difference in the type of

larva (parasitized�non-parasitized) either type of female chose to self-superparsitize,

(x2c1, 0.95�0.52, P�0.05).

Host-searching behavior

Five relevant behaviors were described: walking, host searching, attempts at

oviposition, oviposition, and resting. These behaviors were previously described in

Biosteres longicaudatus (�D. longicaudata) by Lawrence (1981), who considered

only four sequential behaviors: (1) aleatory host searching (behavior that we

Table 1. First choice, number and duration (Mean9SEM) of ovipositions performed by wild

and mass reared D. longicaudata females on parasitized and non-parasitized A. ludens larvae,

during 30-min choice test.

First choice Ovipositions1

Type of female Type of larva Number Number Duration (s)2

Mass reared Parasitized 16 3.390.59 a 50.294.6 a

Non parasitized 9 3.890.37 a 48.292.7 a

Wild Parasitized 8 2.390.33 a 57.596 a

Non parasitized 12 4.090.37 a 6095.8 a

Different letters in the same column indicate significant difference (Ancova, a�0.05).1Mean number and mean duration based on n�8 mass reared females, and n�9 wild females recorded.2Duration includes exploration and oviposition on the same larva.

Biocontrol Science and Technology 141

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considered as walking), (2) Non-aleatory searching (�host searching), (3) ovipositor

insertion, behavior that we separated into attempts of oviposition and oviposition, and

(4) cleaning of ovipositor and antennae at the end of host searching, that we defined

as the resting period.

These behaviors were defined as follows: (1) Walking. The females walked in the

experimental arena with rapid movements. Their antennae were kept parallel to the

substrate with a 458 angle between them. (2) Host searching. The females walked

slowly in the experimental arena, with the antennae touching the substrate at varying

time intervals. During their host searching, females turned in half circles over the

inspected area, making oscillating movements with their ovipositor. (3) Oviposition

attempts (probing): The female inserted her ovipositor into the substrate, but

withdrew it quickly to continue host searching. (4) Oviposition. The female raised

her abdomen and inserted her ovipositor, usually making gentle motions until she

made contact with the host larva. Then, she remained immobile and spread her

antennae to an angle of 180o, which may be coincident with the moment of egg-

laying. Finally, the female made gentle abdominal movements withdrew her

ovipositor and slipped it into its sheath. (5) Resting. Following oviposition, the

females remained motionless, cleaning their antennae and legs, before continuing the

search for more host larvae. Figure 1 shows the flow diagrams for the behavioral

sequences performed by both kinds of females. There were no noticeable differences

between the behavioral sequences of wild and mass-reared females.

Superparasitism by lab and wild females under mass-rearing situation

Both kinds of females tended to superparasitize their hosts, and that super-

parasitism was more intensive (i.e., a higher proportion of exposed larvae was

superparasitized) in direct relation to time (Figure 2; F9, 60�7.90, PB0.001). There

were significant differences between wild and mass-reared females for both

parameters (F1, 60�169.48, PB0.001 for number of scars/puparium, and F1, 60�

59

19

49

24

5026

21

50

59

959

42

4950

21

50

59

9

33

5959

42

49495050

21

31

5050

59

9

WALKING

HOST SEARCHING

OVIPOSITIONATTEMPTS

OVIPOSITION

RESTING

Lab females Wild females

40

36 13

53

23

37

20

39

61

52

44

30

18

4040

36

5353

23

3737

20

39

6161

52

44

WALKING

HOST SEARCHING

OVIPOSITIONATTEMPTS

OVIPOSITION

RESTING

42

a b

Figure 1. Oviposition ethogram of mass-reared (a) and (b) wild Diachasmimorpha long-

icaudata females attacking third-instar Anastrepha ludens larvae. The transition arrows

connect sequential behaviors. The width of the arrows is proportional to the relative transition

frequency. Numbers indicate the observed frequencies.

142 P.I. Gonzalez et al.

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36.16, PB0.001 for proportion of superparasitized hosts). The interaction between

factors (female type and day number) was significant in the case of number of scars

(F4,60�4.76, PB0,0021), but not in the proportion of superparasitized larvae

(F 4,60�0,68, P�0,6071); The superparasitism inflicted by wild females was always

significantly lower. The relationship between the number of scars per puparium and

the proportion of superparasitized larvae was highly significant, but different in

both strains, R2�0.98 for wild females, R2�0.89 for lab females (Figure 3).

Discussion

One of the major findings in this study is the apparent ubiquity of superparasitism in

D. longicaudata females, whether mass reared or field collected. We observed high

Figure 2. Proportion (Mean9SD) of Anastrepha ludens superparasitized larvae (a) and

number of oviposition scars per puparium (b), caused by wild and lab Diachasmimorpha

longicaudata females under mass-rearing conditions. Different letters above bars mean

significant differences between type of female and among days. (a�0.05, PB0.0021 for scars

number; P�0.6071 for proportion of superparasitized larvae).

Biocontrol Science and Technology 143

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levels of superparasitism inflicted by the two strains, both when females singly

exploited a patch (as in Montoya et al. 2000a, 2003) and when other females were

present. However, both strains seemed to select non-parasitized larvae more

frequently after their first oviposition experience. These results suggest that 156

generations under mass-rearing conditions have not significantly influenced the

ovipositional decisions in this species.

Wild and laboratory females also showed similar behavioral patterns in each one

of the phases of oviposition. The observed differences in the behavioral sequences

were mainly related to the intensity of each activity rather than to some variation in

the process of oviposition behavior. According to our data, it seems that the mass-

rearing process has selected for more aggressive, more fertile, and precocious females.

This is reflected in the number of hosts parasitized and the level of superparasitism

observed among mass-reared females under our experimental conditions. A similar

phenomenon has been observed in fruit flies (Liedo and Carey 1996; Miyatake 1998;

Cayol 2000; Liedo, Salgado, Oropeza, and Toledo 2007; Hernandez, Toledo,

Artiaga-Lopez, and Flores 2009), where females become more fertile and precocious

as a consequence of their adaptation to mass-rearing conditions.Ethograms suggested that the behavioral sequences in the choice of either

parasitized larvae or non-parasitized larvae did not differ in wild and lab females.

Foraging behavior was very similar in both strains. There were differences only in the

percentages of transition among behaviors. For example, ‘walking’ was the random

initial behavior prior to host searching for both strains (42% for lab females; 36% for

the wild ones). During this activity, females walked slowly around the experimental

arena, touching the substrate at intervals with their antennae (49% for lab females;

40% for wild females). This activity probably served to locate host larva, although

females also use the mechanoreceptors located in their tarsi for this process (Leyva,

Martınez, and Valdez 1988). According to Lawrence (1981), the duration of this

behavior depends on the ability of the female to orient herself to the body-position of

the host larva inside the fruit.

y = 0,0402x + 0,4784 R2 = 0,8958

y = 0,146x + 0,1339 R2 = 0,9865

0

0,1

0,2

0,3

0,4

0,5

0,6

0,7

0,8

0,9

1

0 2 4 6 8 10

Number of scars / pupa

Pro

port

ion

of s

uper

para

sitiz

ed la

rvae Wild

Laboratory

Figure 3. Relationship between number of scars per puparium and proportion of super-

parasitized larvae in Anastrepha ludens for wild (diamonds) and mass-reared (squares)

Diachasmimorpha longicaudata females.

144 P.I. Gonzalez et al.

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Superparasitism by both wild and lab females under the ratio used in mass rearing

(two host larvae/wasp female), showed that both strains had a similar tendency to

increase the level of superparasitism (i.e., number of oviposition scars per puparium as

in Montoya et al. 2000a) and the proportion of superparasitized larvae over time. This

can be interpreted as a consequence of experience as well as of physiological maturity.

Lawrence, Greany, Nation, and Baranowski (1978) stated that in D. longicaudata

females, the number of mature oocytes tends to increase as experience of ovipositionincreases. The strong tendency of this species to superparasitize even when large

quantities of hosts are available (Montoya et al. 2000a), supports the notion that

superparasitism behavior in D. longicaudata could be an adaptive response. Notably,

adults emerging from host populations with moderate levels of superparasitism

showed the highest percentage of female emergence and adult flyers, without any

deleterious effect in other parameters such as fertility and longevity (Gonzalez et al.

2007). The tendency to superparasitize a host seems to be widespread even among

parasitoid species that show an innate ability for host discrimination (van Lenteren

1981; Tenna, Kapranas, Garcia-Marı, and Luck 2008).

It has been acknowledged that females of solitary parasitoids do not obtain any

benefits from self-superparasitism, except when the total fitness for two immatures in

self-superparasitized hosts is higher than the fitness for a single immature in

parasitized hosts, and when it reduces the risk of further superparasitism by other

females (Waage 1986; Godfray 1994). This improved fitness is probably attributable

to two immatures depressing the host immune system more effectively than one, orincreasing the feeding of the host (Waage 1986; Mackauer and Chau 2001). This may

be the case in D. longicaudata females who introduce entomopoxvirus (DlEPV) into

their hosts during oviposition (Lawrence and Akin 1990). These virus replicate in the

cytoplasm of the host’s hemocytes and inhibit the host’s encapsulation response

(Lawrence 2005). The incidence and intensity of encapsulation decrease as the

number of parasitoid eggs laid in a host increases (including superparasitism;

Blumberg and Luck 1990; Sagarra, Peterkin, Vincent, and Stewart 2000).

The tendency to superparasitize has also been observed in other species of

Hymenoptera under mass-rearing conditions, such as in Trichogramma spp. Smith

(1996) mentioned that a high female: egg host ratio is conducive to superparasitism,

with the adverse consequence of highly male biased offspring sex ratios and low

quality in the insects produced. According to Wanjberg et al. (1989), superparasitism

in T. maidis must be avoided in mass rearing, in order to reduce the risk of low field

efficiency. However, it is important to highlight that the origin of superparasitism in

each species can be due to different factors, such as host availability, a female’s

previous oviposition experience, host physiological resistance, or the adaptivemechanisms exhibited by parasitoids (Vinson 1984; van Alphen and Visser 1990;

Godfray 1994; Rosenheim and Hongkham 1996). Our findings suggest that super-

parasitism must be carefully evaluated in all contexts. Under mass rearing

conditions, it could have adverse consequences or benefits, and influence the success

or failure of biocontrol programs. The prevalence and implications of this

phenomenon under natural field conditions also needs to be investigated. Pre-

liminary observations on D. longicaudata (P. Montoya, unpublished data), indicate

that superparasitism is not unusual in nature.

In summary, our data showed that wild and laboratory reared females possessed

the same basic host discrimination and oviposition behaviors, and that mass rearing

Biocontrol Science and Technology 145

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for 156 generations has only selected females that are more active during host

searching, perhaps because of the historically high densities of competitors. Our

results highlight why mass rearing of this species has been successful and why

D. longicaudata is one of the most effective biological control agents for augmentativereleases against fruit flies (e.g., Sivinski et al. 1996; Montoya et al. 2000b; Ovruski

et al. 2003).

Acknowledgements

We are grateful to John Sivinski (USDA-ARS) for helpful review of this manuscript, Lia Ruizand Patricia Lopez (Moscafrut Program, SAGARPA-IICA) for technical assistance, JavierValle-Mora (ECOSUR) for assistance in statistical analysis and Julio Dominguez (Moscafrutfacility, SAGARPA-IICA) for providing insects. The study received financial support fromCONACYT project no. 185672 to P.G.

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