Biological Control 43 (2007) 188194 www.elsevier.

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Susceptibility of the house y pupal parasitoid Muscidifurax raptor (Hymenoptera: Pteromalidae) to the entomopathogenic bacteria Bacillus thuringiensis and Brevibacillus laterosporus
L. Ruiu *, A. Satta, I. Floris
` Dipartimento di Protezione delle Piante, Sezione di Entomologia agraria, Universita di Sassari, via E. De Nicola, 07100 Sassari, Italy Received 25 February 2007; accepted 20 August 2007 Available online 26 August 2007

Abstract Muscidifurax raptor, an ectophagous pteromalid parasitoid of various muscid ies such as Musca domestica, can be mass-reared and released in poultry and livestock farms for house y control. Entomopathogenic bacteria, such as Bacillus thuringiensis, are commercialized for the biological control of dierent insect pests, but they may have side-eects on other natural enemies. In this study, the susceptibility of M. raptor to three strains of entomopathogenic bacteria (B. thuringiensis subsp. kurstaki (Btk) strain HD1, B. thuringiensis subsp. israelensis (Bti) strain ONR60A, and a new and promising strain of Brevibacillus laterosporus toxic to the house y) was evaluated. Adult wasps were fed a diet treated with high bacterial concentration (>109 spores/g of diet) of Btk, Bti or B. laterosporus. Btk had no detectable eect on M. raptor, B. laterosporus caused only a slight eect on M. raptor adults, and Bti caused a signicant mortality and reduction in the reproductive potential of the wasp. In another test in which parasitoids developed on house y pupae from pre-treated larvae, Bti caused signicant reductions in the development time (6.1%) and in adult emergence rate (up to 59.2%) of M. raptor compared to the control. By contrast, tritrophic interaction (house ybacteriaparasitoid) did not occur with B. laterosporus. The compatibility of the B. laterosporus strain in house y integrated management strategies with parasitoids is promising. 2007 Elsevier Inc. All rights reserved.
Keywords: House y; Muscidifurax raptor; Bacillus thuringiensis; Brevibacillus laterosporus; Microbial control; Pupal parasitoid; Non-target eects

1. Introduction Muscidifurax raptor Girault and Sanders, like other pteromalid wasps, is a microhymenopteran parasitoid of synanthropic and other ies, which plays an important role as a natural biological control agent (Rueda and Axtell, 1985). The possible use of M. raptor in y control programs in poultry and livestock facilities, especially against the house y, Musca domestica L., and the stable y, Stomoxys calcitrans (L.), has been studied in various parts of the world (Olton and Legner, 1975; Rutz and Axtell, 1979). The house y and the stable y are annoyers of medical and veterinary importance and disease vectors of signi-

Corresponding author. Fax: +39 079229329. E-mail address: lucaruiu@uniss.it (L. Ruiu).

cance for humans and animals (Moon, 2002). The widespread use of chemical insecticides against these pests reduces the impact of parasitic groups and promotes the development of insect resistance (Axtell and Arends, 1990). For these reasons, alternative y control methods, such as biological control, are of increasing importance. For instance, the release of lab-reared pteromalids could be employed in integrated management strategies for house y control (Crespo et al., 1998; Hogsette, 1999). Bacillus thuringiensis Berliner is a spore forming bacterium, characterized by its ability to produce parasporal bodies (crystals) which contain specic insecticidal endotoxins. Among entomopathogenic bacteria, it is the most studied species. It is also the most widely used biopesticide in the world (Glare and OCallaghan, 2000). The ecacy of some B. thuringiensis isolates against the house y has been widely demonstrated (Indrasith et al., 1992; Hodgman

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et al., 1993; Johnson et al., 1998; Zhong et al., 2000). On the other hand, more recent studies have focused on lethal or sub-lethal eects of B. thuringiensis on non-target insects (Glare and OCallaghan, 2000). Brevibacillus (formerly Bacillus) laterosporus (Laubach) is an aerobic spore forming entomopathogenic bacterium, characterized by the production of a canoe-shaped lamellar body attached to one side of its spore (Laubach, 1916). Some studies have demonstrated the insecticidal properties of dierent B. laterosporus strains against larvae of mosquitoes (Culex quinquefasciatus Say and Aedes aegypti L.), black ies (Simulium vittatum (Zetterstedt)) (Favret and Yousten, 1985; Rivers et al., 1991), coleoptera (Singer, 1996) and lepidoptera (Oliveira et al., 2004), as well as nematodes and molluscs (Singer, 1996). Very recently a new strain of B. laterosporus showing toxicity against the house y has been isolated (Ruiu et al., 2006). These entomopathogenic bacteria should be compatible with naturally occurring biological control agents, such as hymenopteran parasitoids, in order to maximize biological pest control. However, most experiments have focused on the non-target eects of B. thuringiensis subsp. kurstaki on lepidopteran larval parasitoids (Dunbar and Johnson, 1974; Salama et al., 1991; Atwood et al., 1997; Blumberg et al., 1997) and no data on the susceptibility of parasitoids to the more recently discovered entomopathogen B. laterosporus have been reported so far. In addition, to our knowledge there is very little information about the susceptibility of M. raptor to insecticides (Scott et al., 1991; Geden et al., 1992) and no information is available about the non-target eects of entomopathogenic bacteria on this parasitoid. Our paper deals with laboratory trials investigating the susceptibility of M. raptor to B. thuringiensis subsp. kurstaki strain HD1, widely used for lepidopteran pest control (Glare and OCallaghan, 2000), B. thuringiensis subsp. israelensis strain ONR60A, normally used against mosquitoes and black ies (Goldberg and Margalit, 1977), and the new and promising strain of B. laterosporus known to be highly toxic to the house y (Ruiu et al., 2006). In particular, our work focuses on the toxicity of these three bacterial strains to M. raptor adults by direct ingestion and on the sideeects resulting from the development of immature parasitoids on house y larvae pre-fed with a diet containing bacteria at sub-lethal concentrations. 2. Materials and methods 2.1. Insect rearing Muscidifurax raptor specimens used in the bioassays were reared in our laboratory at the Department of Plant Protection of the University of Sassari. They originated from several house y pupae collected from a cattle farm in Arborea in Central-Western Sardinia (Italy). Adults were maintained in the laboratory at 25 1 C and a photoperiod of L14:D10 in Plexiglas cages

(30 30 30 cm) with two lateral faces covered with gauze to allow ventilation. Honey or sucrose and water (80%) (w/ v) were administered ad libitum as food. Insects were provided with 24- to 48-h-old house y pupae, which after exposure to parasitization, were maintained at 25 1 C to allow development of immature specimens until adult emergence (Petersen, 1986). The house ies employed in M. raptor rearing and bioassays were maintained in our laboratory and came from an original colony supplied by the Entomology Institute of the University of Milan (Italy) and periodically crossed with wild ies. Rearing techniques and conditions were as described elsewhere (Ruiu et al., 2006). 2.2. Bacterial strains and growth conditions Bacillus thuringiensis subsp. kurstaki (Btk) strain HD1 and B. thuringiensis subsp. israelensis (Bti) strain ONR60A were provided by the Bacillus Genetic Stock Center (The Ohio State University, Columbus, Ohio). The B. laterosporus strain used in our study was isolated from a soil sample collected in Sardinia (Italy) and is toxic to the house y (Ruiu et al., 2006). All bacterial strains were grown on nutrient broth, shaken at 30 C until sporulation was complete (Karamanlidou et al., 1991; Ruiu et al., 2006). The suspensions were centrifuged at 11,000g and 4 C for 10 min; the pellets were washed three times in water and adjusted with water to obtain the dierent concentrations of the suspensions used in the bioassays. Presence of spores of B. laterosporus and spores and crystals of B. thuringiensis was conrmed by phase microscopy (1000). After that, concentrations of the dierent bacterial suspensions were measured by Thoma Chamber (E. Hartnack, Berlin, Germany) and expressed as number of spores per gram. 2.3. Lethal bioassays To better evaluate the pathogenic potential of the three studied strains against M. raptor, bacterial concentrations employed were much higher than those that could be used in the eld for insect control. This method permits to determine any unwanted non-target eects of the bacteria, not detectable at lower concentrations (Salama et al., 1991). Over a 7-day period, each bacterial suspension was incorporated at a high concentration into an articial diet and fed to M. raptor adults as follows: four groups of 10 newly emerged adults of both sexes were kept in transparent plastic pots (10-cm diameter 10-cm high) at 25 1 C. They were fed daily with a bacterial suspension that had been diluted 1:1 with a 60% sucrose solution in distilled water to a nal concentration of 2 109 spores per g of diet. The bacterial suspension was placed in a capillary tube and made available to the parasitoids. In the negative control group, insects were fed a 30% sucrose solution. Adult mortality was recorded after 7 days. In addition, to ensure that the insects had fed on the bacterial

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suspension and to demonstrate that they did not die from starvation, an additional control treatment with parasitoids receiving no food was used (Eijs et al., 1998). This experiment was repeated twice. 2.4. Sub-lethal bioassays 2.4.1. Adult parasitoid feeding experiment These bioassays involved two dierent experiments, the rst with B. laterosporus and the second with Bti. Btk was used only in lethal bioassays because it is normally not employed for the control of Diptera (Glare and OCallaghan, 2000). A total of 32 treated parasitoid and 31 control pairs were employed for the experiment with B. laterosporus, whereas 15 treated and 15 control pairs were used in the experiment with Bti. Treated pairs of newly emerged adult parasitoids (1 male and 1 female) were kept in plastic pots (10-cm diameter 10-cm high) and fed daily for 5 days on a liquid diet containing the bacterial suspension at a nal concentration of 1 109 spores/g and 30% sucrose. Control pairs were fed a 30% sucrose solution. Food was administered by capillary tubes as previously described. From the 6th day on, parasitoid pairs which had survived after treatment and control pairs were reared on 30% sucrose solution. Honey was provided ad libitum. Each pot had a window through which each female received 20 house y pupae (24- to 48-h-old) daily until death occurred. Parasitoid mortality was assessed daily until the last one died. After being exposed to parasitization for 1 day, pupae were maintained at 25 C until house y or M. raptor adult emergence. The numbers of emerged parasitoids and their emerging holes on the puparia were recorded. Adult parasitoid emergence rate was then calculated taking into account the number of puparia with an emerging hole and the number of exposed pupae. 2.4.2. Immature house y feeding experiment Groups of ve newly emerged and just mated M. raptor females were kept in plastic pots (10-cm diameter 10-cm high) and provided with 10 house y pupae (24-h-old) developed from larvae treated with either Bti or B. laterosporus. These pupae were from either 1st or 2nd instar larvae reared on a diet containing a bacterial concentration of 107 or 108 spores/g, respectively. Because of their higher susceptibility, younger house y larvae were exposed to a lower bacterial concentration to ensure their development and pupation (Ruiu et al., 2006). In control groups, pupae were from non-treated larvae. After being exposed to parasitization for 3 h, house y pupae were maintained at 25 C until adult emergence. Each treatment had three replicates in the rst trial and six replicates in the second trial. Adult parasitoid emergence rate was calculated as in the previous experiment. Emerged parasitoid adults were kept in plastic pots (one for each replicate) and fed honey and water ad libitum.

Adult survival was recorded daily and longevity was then calculated. 2.5. Statistical analysis In the case of lethal bioassays and in the immature house y feeding experiment (sub-lethal bioassays), data on mortality percentages of parasitoid adults, immature development time, parasitoid emergence rate and adult longevity, from both trials, were combined for analysis and presentation because variance analyses for each of the two experiment trials gave similar results. These data were analyzed as a two factor design (ANOVA: treatment, experiment repetition) followed by an LSD test to separate treatment means, using Statgraphics Plus (2001) software. In the adult parasitoid feeding experiments (sub-lethal bioassays), data on longevity, number of emerged adults per female and percentages of emergence were analyzed using t-tests to compare means of treated and control groups. In all experiments, data met the assumption of homogeneity of variance. 3. Results 3.1. Lethal bioassays Mortality percentages of M. raptor adults caused by the dierent bacterial suspensions at the concentration of 2 109 spores/g of diet for 7 days were signicantly aected by treatment (F = 24.21; df = 3, 24; P < 0.0001), whereas there was no eect of experiment repetition (F = 0.51; df = 1, 24; P = 0.4823) and no interaction between treatment and experiment repetition (F = 0.06; df = 3,24; P = 0.9819). The percentage of mortality (means SEM) caused by B. laterosporus (33.8 2.6%) and Bti (40.0 3.3%) diered signicantly from the negative control (13.8 1.8%). By contrast, no signicant lethal eects were associated with Btk strain HD1 (16.3 1.8%). In the additional control constituted by wasps which received no food, mortality reached 100% after 3 days. 3.2. Sub-lethal bioassays 3.2.1. Adult parasitoid feeding experiment Data on longevity of M. raptor adults fed diets treated with B. laterosporus or Bti at 1 109 spores/g of diet are presented in Table 1. Longevity of M. raptor females fed B. laterosporus spores was signicantly lower (10.4%) than that of the control (t = 2.1716, df = 61, P = 0.0337). The reduction in male longevity caused by the same treatment, however, was not statistically signicant (t = 1.84526, df = 61, P = 0.0698). Similarly, Bti treatments caused a signicant reduction (30.3%) in parasitoid female longevity (t = 5.8481, df = 28, P < 0.00001), whereas male longevity was not aected (t = 1.4584, df = 28, P = 0.1559).

L. Ruiu et al. / Biological Control 43 (2007) 188194 Table 1 Longevitya (means SEM) of Muscidifurax raptor adults treated with Brevibacillus laterosporus or Bacillus thuringiensis subsp. israelensis Experiment Treatmentb Longevity Males 1 2
a

191

Females
c

B. laterosporus Control Bt. subsp. israelensis Control

15.4 0.6a 17.4 0.9a 15.5 1.0a 17.6 1.1a

17.3 0.6a 19.3 0.7b 13.6 0.6a 19.5 0.8b

The longevity is expressed in days. The bacterial suspension was administered at a concentration of 1 109 spores/g of diet. c For each experiment, means in each column followed by dierent letters are signicantly dierent (t-test, P < 0.05).

Data on the emergence of adult parasitoids from house y pupae parasitized by females treated with B. laterosporus or Bti are presented in Table 2. In both studies, the total number of exposed pupae in the treated group was lower than in the control, as a consequence of the shorter lifespan of the treated females. Levels of parasitoid emergence were similar in the B. laterosporus treated group and in the control (t = 0.0984443, df = 61, P = 0.9219). Even though the mean number of emerged adults per ovipositing female was reduced (16.3%) by B. laterosporus treatment, the dierences between treated and control groups were not signicant (t = 1.4817, df = 61, P = 0.1436). When Bti was added to the adult diet, a signicant decrease (25.9%) in the emergence of parasitoids from parasitized pupae was observed compared to that of the control group (t = 3.0544, df = 28, P = 0.0049). Similarly, a signicant reduction (56.4%) in the mean number of emerged adults per ovipositing female was caused by Bti (t = 5.9241, df = 28, P < 0.00001). 3.2.2. Immature house y feeding experiment The treatments of immature house ies with Bti or B. laterosporus did not cause any signicant eect on the development time of immature M. raptor males (treatments on 1st instar larvae: F = 0.28; df = 2, 18; P = 0.7572; treatments on 2nd instar larvae: F = 1.07; df = 2, 18; P = 0.3646) and neither experiment repetition eects (1st

instar: F = 0.41; df = 1, 18; P = 0.5277; 2nd instar: F = 0.03; df = 1, 18; P = 0.8644), nor interaction between experiment repetition and treatment (1st instar: F = 0.57; df = 2, 18; P = 0.5771; 2nd instar: F = 0.22, df = 2, 18; P = 0.8081) were observed (Table 3). Similarly, these two treatments had no eects on the development time of M. raptor females in house y pupae from treated 1st instar larvae (F = 0.72, df = 2, 17; P = 0.4996) and both experiment repetition eects (F = 0.64, df = 2, 17; P = 0.4336) and the interaction between experiment repetition and treatment (F = 0.01, df = 2, 17; P = 0.9896) were not signicant (Table 3). By contrast, when 2nd instar house y larvae where reared on a diet treated with Bti, a slight but statistically signicant reduction (6.1%) in immature female parasitoid development time, compared to B. laterosporus and control groups, was determined (F = 11.61, df = 2, 22; P = 0.0004). On the other hand, the inuence of experiment repetition (F = 0.21, df = 1, 22; P = 0.6539) and the interaction between experiment repetition and treatment (F = 0.67, df = 2, 22; P = 0.5209) were not signicant (Table 3). M. raptor emergence from house y pupae exposed to parasitization was signicantly reduced, when pupae developed from 1st or 2nd instar larvae reared on a diet containing Bti (1st instar: F = 11.68; df = 2, 21; P = 0.0004; 2nd instar: F = 10.42; df = 2, 21; P = 0.0007). In comparison to the control group, the reduction in parasitoid emergence was of 59.2% and 56.1% when 1st instar and 2nd instar house y larvae, respectively, were treated with Bti (Table 4). In contrast, B. laterosporus did not signicantly aect adult emergence of M. raptor. Neither experiment repetition eects (1st instar: F = 0.28; df = 1, 21; P = 0.6011; 2nd instar: F = 0.01, df = 1, 21 P = 0.9999) nor interaction between experiment repetition and treatment (1st instar: F = 0.05; df = 2, 21; P = 0.9560; 2nd instar: F = 0.09, df = 2, 21; P = 0.9116) were noticed. Longevity of adults emerging from these pupae was not inuenced by treatments on 1st instar (male: F = 0.32; df = 2, 52; P = 0.7267; female: F = 0.23; df = 2, 39; P = 0.7949) or 2nd instar (male: F = 0.04, df = 2, 49; P = 0.9602; female: F = 0.04, df = 2, 38; P = 0.9602) house y larvae (Table 5). Similarly, there were no signicant eects of experiment repetition (1st instar: F = 0.01;

Table 2 Emergence of Muscidifurax raptor adults from house y pupae parasitized by females treated with Brevibacillus laterosporus or Bacillus thuringiensis subsp. israelensis Experiment 1 2 Treatmenta B. laterosporus Control Bt. subsp. israelensis Control
a b

Ovipositing females (n) 32 31 15 15

Exposed pupae (n) 7080 8300 2280 4120

Emerged adults (n) 1941 2249 487 1117

Emerged adults per female (means SEM) 60.7 5.7ab 72.5 5.7a 32.5 4.1a 74.5 5.8b

Percentage of emergence (means SEM) 26.5 1.6a 26.7 1.3a 20.9 1.5a 28.2 1.9b

The bacterial suspension was administered at a concentration of 1 10 spores/g of diet. For each experiment, means in each column followed by dierent letters, are signicantly dierent (t-test, P < 0.05).

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Table 3 Development timea (means SEM) of Muscidifurax raptor females and males on house y pupae from 1st and 2nd instar larvae treated with dierent concentrations of Brevibacillus laterosporus or Bacillus thuringiensis subsp. israelensis Treatmentb House y larval instar 1st Males Bt. subsp. israelensis B. laterosporus Control
a b

P = 0.9797 for males and females, respectively) (2nd instar: F = 0.01; df = 2, 49; P = 0.9856 and F = 0.09; df = 2, 38; P = 0.9151, for males and females, respectively). 4. Discussion

2nd Females 24.6 0.2a 24.6 0.1a 24.4 0.1a Males 21.8 0.2a 21.4 0.1a 21.6 0.1a Females 23.2 0.3a 24.4 0.2b 24.7 0.2b

21.1 0.2ac 21.3 0.1a 21.3 0.2a

The development time is expressed in days. 1st and 2nd instar housey larvae were reared on diets containing a bacterial concentration of 1 107 and 1 108 spores/g of diet, respectively. c Means in each column followed by dierent letters are signicantly dierent (LSD test, P < 0.05).

Table 4 Numbers (means SEM) of Muscidifurax raptor emerged from 10 house y pupaea from 1st and 2nd instar larvae treated with dierent concentrations of Brevibacillus laterosporus or Bacillus thuringiensis subsp. israelensis Treatmentb House y larval instar 1st Bt. subsp. israelensis B. laterosporus Control
a b

2nd
c

2.0 0.2a 4.6 0.4b 4.9 0.5b

1.8 0.4a 4.8 0.4b 4.1 0.5b

Ten house y pupae were exposed to ve M. raptor females for 3 h. 1st and 2nd instar housey larvae were reared on diets containing a bacterial concentration of 1 107 and 1 108 spores/g of diet, respectively. c Means in each column followed by dierent letters are signicantly dierent (LSD test, P < 0.05).

Table 5 Longevitya (means SEM) of Muscidifurax raptor females and males developed on house y pupae from 1st and 2nd instar larvae treated with dierent concentrations of Brevibacillus laterosporus or Bacillus thuringiensis subsp. israelensis Treatmentb House y larval instar 1st Males Bt. subsp. israelensis B. laterosporus Control
a b

2nd Females 20.5 0.9 21.1 0.7 20.2 0.9 Males 17.6 1.1 17.7 0.6 17.9 0.7 Females 20.7 0.6 20.8 0.7 21.1 0.7

16.5 1.0c 17.5 0.7 17.8 0.6

Longevity is expressed in days. 1st and 2nd instar housey larvae were reared on diets containing a bacterial concentration of 1 107 and 1 108 spores/g of diet, respectively. c Means in each column are not signicantly dierent (ANOVA, P > 0.05).

df = 1, 52; P = 0.9634 and F = 0.01, df = 1, 39; P = 0.9621 for males and females, respectively) (2nd instar: F = 0.09; df = 1, 49; P = 0.7707 and F = 0.01, df = 1, 38 P = 0.9613, for males and females, respectively) and no interaction between treatment and experiment repetition (1st instar: F = 0.32; df = 2, 52; P = 0.7255 and F = 0.02; df = 2, 39;

In general, B. laterosporus and Bti exhibited weak lethal eects on M. raptor when administered with food at concentration levels to which the house y and mosquitoes are known to be highly susceptible (Goldberg and Margalit, 1977; Favret and Yousten, 1985; Rivers et al., 1991; Ruiu et al., 2006). The pteromalid was more sensitive to Bti than to B. laterosporus and was not susceptible to Btk. In natural conditions, food uptake behavior of pupal parasitoids may limit direct contacts with entomopathogenic bacteria applied in a eld. However, the host-feeding behavior of M. raptor adults (Legner and Gerling, 1967) and the development of their immature stages on y pupae may be a way of indirect contact with bacteria or their residue (fresh or dry). On the other hand, M. raptor adults might be aected by low concentrations of a bacterial insecticide by feeding on contaminated material or on treated insect hosts. For these reasons, sub-lethal eects of these bacteria should be considered when evaluating their potential eects on non-target insects such as M. raptor. When adult wasps were fed a diet treated with Bti, the observed reductions in female longevity and emergence rate of the following generation demonstrated a decrease in the reproductive potential of M. raptor. This might have been partly due to the reduction in female lifespan. By contrast, the only secondary eect of feeding M. raptor adults diets treated with B. laterosporus was a slight reduction in female longevity. Considering the high concentrations assayed, the observed eects seem to be due mainly to a general pathogenicity rather than to a specic toxicity of Bti to this insect species. After all, it is known that nutrition and oviposition of newly emerged M. raptor females inuence their subsequent behavior and reproductivity (Legner and Gerling, 1967). Some studies on diverse hymenopteran parasitoid species showed a reduction in the longevity and parasitization activity of adults fed a diet containing B. thuringiensis (Dunbar and Johnson, 1974; Salama et al., 1991). In other cases, however, feeding a diet treated with that bacterium was not harmful to adult parasitoids (Blumberg et al., 1997). Rearing M. raptor on M. domestica larvae pre-fed with Bti reduced the development time of the parasitoid and, consequently, the adult emergence rate as well. The fact that such eects were more noticeable when immature wasps developed on house y pupae from pre-treated 2nd instar larvae might be related to the higher bacterial concentration to which these larvae were exposed. Similarly, Salama et al. (1991) showed that development and emergence of the braconid, Bracon brevicornis Wesm., were negatively aected when female parasitized Plodia

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interpunctella Hubner larvae pre-fed a diet containing Btk HD1. Similar eects were previously reported by El-Maghraby et al. (1988) on Microplitis ruventris Kok. parasitizing B. thuringiensis-treated Spodoptera littoralis (Boisd.) larvae. In addition, Blumberg et al. (1997) reported that pre-feeding Helicoverpa armigera (Hu bner) larvae with Btk HD1 prevented the successful development of its larval endoparasitoid, the braconid Microplitis croceipes (Cresson). Oviposition and development of M. raptor did not seem to be aected in pupae of M. domestica developed from larvae pre-fed with B. laterosporus. Although pteromalid development is known to be inuenced by the quality of food (Coats, 1976), these ndings suggest that wasp larvae are not aected by the possible weight loss or the increased development time of immature ies intoxicated with this bacterium (Ruiu et al., 2006). In general, the ability of this parasitoid to develop on pre-treated immature ies avoids any waste of eggs, thus not aecting its reproductive potential. A delayed development of treated house ies that would cause a longer pupal stage in the eld is desirable to favor parasitization (Weseloh and Andreadis, 1982). In conclusion, M. raptor is slightly susceptible to Bti when feeding on a treated diet or developing on pre-treated immature ies. By contrast, the parasitoid adults are susceptible only to high concentrations of B. laterosporus and, apparently, there is no tritrophic interaction (house yparasitoidbacteria) when parasitoids develop on pupae from pre-treated house y larvae. Based on our ndings that the bacterial concentrations parasitoids may come into contact in the eld are obviously lower than those assayed in our laboratory study, the use of B. laterosporus against M. domestica is promising. Although further studies are needed to detect any possible non-target eects of the tested strain on other natural enemies (parasitoids and predators), pteromalid wasps are the main house y parasitoids and M. raptor is one of its most represented species around the world (Rueda and Axtell, 1985). For these reasons, the new strain of B. laterosporus is considered a selective microbe with potential use as a microbial agent for house y management. Acknowledgments This study was supported by a grant from the Italian ` Ministero Universita Ricerca Scientica (Research Program: Biotecnologie innovative per il controllo di insetti nocivi mediante limpiego di agenti microbiologici. Coordinator: Prof. Ignazio Floris, University of SassariItaly). The authors are grateful to Dr Ana H.D. Francesconi for her assistance. References
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